超敏蛋白对烟草黄瓜花叶病毒的诱导抗性及生长、品质的影响

为明确超敏蛋白对烟草黄瓜花叶病毒(Cucumber mosaic virus,CMV)的诱导抗性及其生长、品质的影响,采用半叶枯斑法和田间试验研究了10%超敏蛋白对枯斑三生烟和普通烟的诱导抗性及促生作用,并分析了对烤烟经济性状和内在质量的影响。结果表明,10%超敏蛋白在活体外对CMV无钝化作用,但预防、治疗效果显著,接种前48 h喷施预防效果达75.70%,接种后24 h喷施治疗效果达65.45%,均显著高于对照;10%超敏蛋白处理后烟草的株高、茎围、最大叶面积均高于其它处理,单位产量比20%吗啉胍·乙铜可湿性粉剂处理增产211.8 kg/hm2,增值3 519.2元/hm2;经10%超敏蛋白处理后的烤烟化学成分均处于优质烟的适宜质量分数范围内。表明超敏蛋白不仅有利于提高烟草抗性和促进烟株生长,且可以明显提高烟叶经济效益和烟叶内在质量。     

A Survey of Viruses of Alstroemeria in the UK and the Characterisation of Carlaviruses Infecting Alstroemeria

Alstroemeria samples collected in the UK were tested for a range of viruses using ELISA. Alstroemeria mosaic virus (AlMV), alstroemeria carlavirus (AlCV), lily symptomless virus (LSV), cucumber mosaic virus (CMV) and tobacco rattle virus (TRV) were detected either singly or in combination in 67.5% of 203 samples. AlCV and LSV isolates from Alstroemeria and lily were studied and characterised serologically using existing antisera, and by PCR, using primers to an 11kDa open reading frame (ORF) unique to carlaviruses and to the coat protein gene of LSV. Sequences of isolates of AlCV and LSV from the coat protein gene were 94–99% similar and were 99% similar in the 11kDa ORF, supporting the view that these are strains of the same virus.     

Ornamental plants and thrips populations associated with tomato spotted wilt virus in Greece

A survey was conducted in order to record the ornamental plants that are hosts of tomato spotted wilt virus (TSWV) and impatiens necrotic spot virus (INSV) in Greece. Polyclonal antibodies prepared against the N protein of a Greek isolate of TSWV fromGerbera jamesonii (GR-34) were used. Leaf samples were taken from plants showing typical symptoms of tospovirus infection such as chlorotic and necrotic rings on the leaves and malformation and necrosis of the flowers. The samples were tested by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using polyclonal antibodies to the N proteins of TSWV and INSV (NL-07). ELIS A-positive samples were mechanically transmitted to plants ofPetunia hybrida, Nicotiana rustica andN. benthamiana to confirm infection. Although none of the samples was found infected with INSV, TSWV presence was recorded in 42 botanical species that belong to 40 genera in 27 families. Among them the speciesBeloperone guttata, Coleus barbatus, Impatiens petersiana andLilium auratum are reported for the first time as hosts of TSWV, whereasBegonia sp.,Catharanthus roseus Celosia cristata, Dianthus chinensis, Fuchsia hybrida andStephanotis floribunda are found as new hosts of the virus in Greece. Thrips collected from TSWV-infected plants were in most cases identified asFrankliniella occidentalis, except from plants ofDendranthema sp. andDianthus caryophyllus whereThrips tabaci individuals were also identified. Different percentages of transmitters were noticed when the thrips populations collected from TSWV-infected ornamental hosts were tested for transmission of TSWV.     

Endophytic fungi and ergot alkaloids in native Turkish grasses

Mature inflorescences of 76 samples of ten genera of native grasses were collected in the Ankara province of Turkey. Microscopic examination of individual seeds revealed the presence of endophytic fungi in four seed samples, The endophytes were isolated in pure culture and identified. Seeds were also ground and subjected to ELISA and HPLC analyses to confirm the levels of ergot alkaloids typically produced by endophytic fungi of grasses.     

四种杀菌剂对烟草灰霉病菌的毒力及对烟草灰霉病的抑制作用

为筛选防治烟草灰霉病的有效药剂,采用菌丝生长速率法和孢子萌发法测定氟啶胺、咪鲜胺、苯醚甲环唑及代森锰锌4种杀菌剂对烟草灰霉病菌Botrytis cinerea的毒力,同时通过离体叶片评价这4种杀菌剂对烟草灰霉病的保护和治疗作用。结果表明,氟啶胺和咪鲜胺对菌丝生长活性抑制最强,有效抑制中浓度ECjs50平均值分别为0.02、0.03 mg/L,苯醚甲环唑次之,代森锰锌最弱,EC_(js50)平均值分别为0.39、7.86 mg/L;氟啶胺对孢子萌发活性抑制最强,代森锰锌次之,有效抑制中浓度EC_(mf50)平均值分别为0.06、0.16 mg/L,咪鲜胺和苯醚甲环唑最弱,ECmf50平均值均大于25.00 mg/L。离体试验表明,氟啶胺对烟草灰霉病保护作用最强,浓度为50 mg/L时,防治效果为100.00%,咪鲜胺和苯醚甲环唑次之,防治效果分别为88.62%和76.46%,代森锰锌最弱,浓度为1 000 mg/L时防治效果仅为75.81%;氟啶胺对烟草灰霉病治疗作用最强,浓度为100 mg/L时的防治效果为85.75%,咪鲜胺和苯醚甲环唑次之,浓度为200 mg/L时的防治效果分别为85.47%和76.48%,代森锰锌最弱,浓度为1 000 mg/L时防治效果为70.24%。表明氟啶胺和咪鲜胺更适合烟草灰霉病的防治。     

山东省樱桃主产区果蝇的调查与鉴定

为了解山东省樱桃主产区果蝇的发生情况,本研究选择了山东省主要樱桃产区的7个地点进行调查,并通过形态学特征和分子鉴定法对果蝇的种类进行鉴定。结果发现,山东省樱桃主产区有4种果蝇,通过形态特征与分子鉴定法最终确定4种果蝇,分别为斑翅果蝇Drosophila suzukii、黑腹果蝇D.melanogaster、海德果蝇D.hydei和伊米果蝇D.immigrans。这4种果蝇对樱桃的为害程度与种植地区、果实腐烂程度和樱桃的品种均有关系;莱芜和泰安地区果蝇分别有4种和3种;烟台和威海地区则只有斑翅果蝇和黑腹果蝇2种。烟台地区果蝇为害率最高,且数量最多,其中烟台丁家夼村樱桃为害率高达45.16%,在第2次采集中烟台薛家村每100粒樱桃中斑翅果蝇和黑腹果蝇的数量最多。美早和先锋2个品种樱桃对果蝇的吸引力较强;采摘后期腐烂的果实可吸引大量的果蝇产卵。因此,应对烟台地区进行重点预防、早期防治,并减少美早和先锋品种樱桃在山东省的种植。     

感染马铃薯Y病毒烟株对烟蚜取食行为及烟蚜茧蜂适应性的影响

为探明烟蚜茧蜂Aphidius gifuensis Ashmead对烟蚜Myzus persicae(Sulzer)取食感染马铃薯Y病毒(Potato virus Y,PVY)烟株的适应性,利用刺探电位图谱(EPG)技术记录了烟蚜在感病烟株与健康烟株上的取食行为,并测定了烟蚜茧蜂对取食2种烟株烟蚜的寄生率以及烟蚜茧蜂的羽化率、发育时间及性比等指标。结果表明,与健康烟株相比,烟蚜在感染PVY烟株上的第1次刺探取食持续时间显著延长,且口针遇到阻力的次数(F波)和总持续时间均显著减少。烟蚜在感病烟株木质部的吸食时间(G波)显著长于健康烟株。感病烟株上烟蚜在韧皮部阶段的分泌唾液时间(E1波)较在健康烟株上显著缩短,而被动吸食汁液时间(E2波)显著延长。烟蚜茧蜂寄生取食感染PVY烟株的烟蚜,虽然能成功完成其生活史,但适应性与寄生取食健康烟株烟蚜的蚜茧蜂存在差异。在感染PVY烟株上,烟蚜茧蜂对烟蚜的寄生率为33.67%,显著低于对照的64.67%;且僵蚜体重明显下降,羽化的成蜂个体较小;成蜂存活时间1.48 d也极显著短于对照组的2.25 d。表明烟蚜茧蜂对取食感染PVY烟株烟蚜的适应性较低,PVY可通过烟蚜为介体间接降低烟蚜茧蜂的适应力。     

A survey of the Eriococcidae (Hemiptera: Coccoidea) of Ankara Province,Turkey

The Coccoidea of cultivated and non-cultivated areas of Ankara Province, in central Anatolia, Turkey, were surveyed between 1999 and 2001. Eleven species of Eriococcidae were collected, of which nine are new records for Turkey:Acanthococcus greeni Newstead,A. micracanthus Danzig,A. munroi Boratynski,A. roboris (Goux),A. zernae Tereznikova,Rhizococcus cingulatus (Kiritchenko),R. cynodontis (Kiritchenko),R. herbaceus Danzig andR. pseudinsignis (Green). http://www.phytoparasitica.org posting Sept. 30, 2003.     

黄瓜花叶病毒和马铃薯Y病毒混合侵染烟株对烟蚜取食行为的影响

为探究黄瓜花叶病毒(Cucumber mosaic virus,CMV)、马铃薯Y病毒(Potato virus Y,PVY)混合侵染烟株对烟蚜取食行为的影响,利用刺探电位图谱(electrical penetration graph,EPG)技术记录了烟蚜在健康烟株与CMV、PVY混合侵染后不同发病级别烟株上的取食波形。结果显示:烟蚜在健康烟株上的刺探次数最少,在感病烟株上的C波总持续时间均显著长于健康烟株;第1次到达韧皮部前的刺探次数,健康植株上仅为4.00次,3级感病烟株上的为健康烟株上的2倍;在健康烟株上E2波总持续时间为120.65 min,极显著大于2级和3级感病烟株;刺探过程中,感病烟株上的pd波出现次数均高于健康烟株,且pd波II-1和II-3亚波的持续时间也显著高于健康烟株。研究表明,CMV、PVY混合侵染烟株可降低寄主对烟蚜的适合度,且能促进烟蚜对病毒的传播。     

Choanephora blight of potatoes and other crops grown under tropical conditions in Peru

Samenvatting In het kader van een veredelingsprogramma voor de aanpassing van de aardappel aan de teelt in het warme, vochtige tropische laagland worden sinds 1975 aardappels verbouwd op een proefveld te Yurimáguas in het Amazonebekken van Peru. Vanaf het begin is veelvuldig een ernstige aantasting van het loof waargenomen veroorzaakt doorChoanephora cucurbitarum en enkele nauwverwante soorten. Het ziektebeeld is gekenmerkt door lesies die zich aanvankelijk van bladranden en bladspitsen uitbreiden, waarbij aangetaste weefsels eerst waterig worden en daarna uitdrogen en een olijfgroene kleur aannemen. Bij droog weer worden vier tot vijf mm lange sporangi?ndragers gevormd op de aangetaste delen en ontstaat het voor de ziekte zo kenmerkende beeld (Fig. 2). Dezelfde schimmel is ook gevonden op andere cultuurgewassen (Fig. 1) en is ter plaatse waarschijnlijk de meest beperkende factor voor vele gewassen gedurende regenrijke perioden.     

Yellows of lettuce and some other vegetable crops in the Netherlands caused by beet western yellows virus

A virus isolated from lettuce (Lactuca sativa), endive (Cichorium endivia), witloof chicory (C. intybus), and spinach (Spinacia oleracea), and from some weeds was shown to be beet western yellows virus (BWYV) by its host range, particle morphology and serology. It resembled previously described European isolates but differed from American strains in its inability to infectBeta vulgaris, Brassica pekinensis andRaphanus sativus. The most useful host for routine indexing wasCrambe abyssinica. Virus particles in purified preparations stained with uranyl acetate were isometric, ca. 27 nm in diameter. Purified virus reacted with antiserum to an American strain of BWYV in infectivity neutralization gel diffusion and serologically specific electron-microscopy tests.The field reaction to BWYV of cultivars of lettuce, otherLactuca species and someCichorium species was investigated and differences in symptom expression were observed. On the basis of observations during two seasons BWYV appeared to be widely distributed but seemed of minor economic importance to lettuce growing. It may be a potentially important pathogen of endive and chicory.Samenvatting Reeds gedurende enkele jaren trekt in Nederland een vergelingsziekte van sla (Fig. 1 en 2) de aandacht. In 1977 en 1978 werd de ziekte nader bestudeerd en ook waargenomen in andijvie, witlof (Fig. 3) en spinazie. Uit zieke planten van deze vier gewassen en uit de onkruiden herderstasje en kruiskruid, groeiend in de buurt van de zieke sla, kon doorMyzus persicae op persistente wijze een virus worden overgebracht. Het werd op grond van zijn waardplantenreeks (Tabel 1), deeltjesmorfologie (Fig. 4A) en serologie (Fig. 4B) herkend als het in de USA beschreven beet western yellows virus (BWYV).Het Nederlandse virus komt overeen met in andere landen gerapporteerde Europese isolaten van het virus, maar verschilt van Amerikaanse doordat het niet in staat is om biet, chinese kool en radijs te infecteren. Daarom is voor het virus door Bos en Ashby (1978) de Nederlandse naam slavergelingsvirus ingevoerd. De meest geschikte indicatorplant voor routinetoetsing isCramble abyssinica (Tabel 2). De reactie van herderstasje varieert al naar individu van nagenoeg letaal tot vrijwel symptoomloos (Fig. 5).In gedeeltelijk gezuiverde preparaten bleken de deeltjes bolvormig te zijn en ca. 27 nm in diameter (Fig. 4A). Zulke preparaten reageerden met antiserum tegen een Amerikaanse stam van het virus (BWYV) in toetsen die gebruik maken van infectieneutralisering, gel-diffusie en serologisch-speciefieke elektronenmikroskopie. Bij laatstgenoemde techniek werd een fraaie deeltjesklontering waargenomen (Fig. 4B), die ontbrak na incubatie van gezuiverd virus met een antiserum tegen het niet verwante kersebladrolvirus (Fig. 4C).Bij veldwaarneming in 1977 van 20 slarassen, waarbij tot 60% van de planten van één ras werden aangetast, bleken twee rassen niet of weinig vatbaar (Tabel 3). Bij inoculatie in de kas bleken ze echter volledig vatbaar. In 1978 werd een aantal soorten en rassen vanCichorium enLactuca blootgesteld aan natuurlijke en aan kunstmatige infectie. BehalveC. intybus Groenlof IVT waren allen vatbaar, ookL. sativa Gallega de Invierno,L. serriola enL. virosa.Het virus lijkt algemeen voor te komen. Meestal is de infectiegraad niet hoog. Vanwege de lange incubatieduur in sla is het virus in dat gewas bij de hier toegepaste teeltwijze waarschijnlijk van geringe betekenis. Het lijkt echter een potentieel belangrijk pathogeen voor andijvie en witlof.Guest research worker, Plant Diseases Division, DSIR, Private Bag, Christchurch, New Zealand, with financial assistance from the International Agricultural Centre, Wageningen, and Ministry of Education and Science, The Hague.     

烟草叶斑病病原菌的鉴定及其生物学特性

为明确贵州省新发现的烟草叶斑病病原菌种类和生物学特性,2019年自田间采集具有典型发病症状的烟叶,采用组织分离法对其进行分离纯化,对分离菌株进行致病性测定,观察病原菌形态学特征,基于ITS、LSU、tub2和rpb2基因序列进行分子生物学鉴定,同时对病原菌的生物学特性进行测定。结果显示,烟草叶斑病病斑呈圆形或椭圆形,病斑中间呈浅褐色,边缘呈棕色,周围环绕黄色晕圈。经分离纯化共获得3株病原菌,形成的菌落为深褐色,气生菌丝呈白色;分子孢子器呈球形或扁球形,为深褐色,大小为79.3～91.7μm×148.4～167.3μm;分生孢子呈椭圆形,光滑,无隔膜,具0～3个油球,大小为3.5～5.6μm×1.7～2.9μm。根据形态学特征和分子生物学特征将其鉴定为Didymella segeticola。该病原菌菌丝生长适宜温度为20～25℃;适宜pH为6～10,最适碳源为乳糖,最适氮源为蛋白胨,通气和光暗交替环境有利于其生长。     

嘧菌酯与吡唑醚菌酯对烟草白粉病菌的抑制作用及在烤烟漂浮育苗阶段的防治效果

采用小苗法测定了嘧菌酯和吡唑醚菌酯对烟草白粉病菌Erysiphe cichoracearum的毒力,以及对苗期烟草白粉病的预防和治疗作用及持效期。结果表明:嘧菌酯和吡唑醚菌酯对烟草白粉病菌抑制作用的EC50值分别为7.62和9.90 μg/mL,对该病害的预防和治疗作用的EC50值分别为8.76、12.34 μg/mL和17.32、23.40 μg/mL;2种药剂对烟草白粉病菌均具有较长的持效期,在有效成分56.25 g/hm2 剂量下、2次喷雾处理烟苗,30 d后的防效仍分别达到94.45%和88.53%;2种药剂对新生烟叶均具有良好的保护作用,且在实验剂量范围内对烟苗安全。     

Plant Host Range of Verticillium longisporum and Microsclerotia Density in Swedish Soils

Verticillium longisporum is a soil-borne fungal pathogen causing vascular wilt of Brassica crops. This study was conducted to enhance our knowledge on the host range of V. longisporum. Seven crop species (barley, oat, oilseed rape, pea, red clover, sugar beet and wheat) and five weed species (barren brome, black-grass, charlock, cleavers and scentless mayweed) all common in southern Sweden were evaluated for infection by response to V. longisporum. Oat, spring wheat, oilseed rape, scentless mayweed and charlock inoculated with V. longisporum in a greenhouse showed stunting to various degrees close to the fully ripe stage. Based on the extent of microsclerotia formation, explants were separated into four groups: for pea and wheat, <5% of the samples had formed microsclerotia; for scentless mayweed, 5–10%; for oat, 10–20%; and for charlock and oilseed rape >80%. The results suggest that plant species outside the Brassicaceae can act as reservoirs of V. longisporum inoculum. Soil inoculum densities in nine fields were monitored over a period of 12 months, which ranged from 1 to 48 cfu g⁻¹ soil. Density of microsclerotia was lowest just after harvest, reaching its maximum six months later. No significant correlation between inoculum density in soil and disease incidence on oilseed rape plants was found. However, the data suggest that a threshold of 1 cfu g⁻¹ soil is needed to cause disease on oilseed rape. Species identification based on microsclerotia morphology and PCR analysis showed that V. longisporum dominated in soil of seven, and V. dahliae in two of the nine fields studied.     

烟草青枯病菌在烟草根际的定殖及最适发病条件

为明确烟草青枯病菌在根际的定殖情况和最适发病条件,采用绿色荧光蛋白(gfp)标记病原菌,监测其在根际的定殖位置及数量动态变化,并运用荧光定量PCR法测定病原菌在根际和土体土壤的数量,通过响应曲面法探索病原菌致病的临界浓度、发病最适温湿度以及对烟株青枯病发病的影响。结果表明,从贵州省烟田土壤中分离到烟草青枯病病原菌茄科劳尔氏菌Ralstonia solanacearum,gfp-标记的茄科尔劳尔氏菌主要定殖于根尖和根毛区,少数菌落定殖于初生根的延长区,定殖部分表现为非连续性;第10、15、20天根际土壤的茄科劳尔氏菌数量均显著高于土体土壤数量,分别是土体土壤中的1.15、1.33、1.42倍;当土壤中每克土病原菌数量为106.82 CFU、温度为30.55℃、相对湿度为81.42%以上时,烟草青枯病的病情指数最高为91.13,烟株易发生青枯病。表明茄科尔劳尔氏菌在根际土壤中比土体土壤更易定殖,烟草青枯病发病条件主要取决于病原菌浓度、温度以及相对湿度3项指标。     

Immunochromatographic assay for simple and rapid detection of <Emphasis Type="Italic">Satsuma dwarf virus</Emphasis> and related viruses using monoclonal antibodies

A simple and rapid immunochromatographic assay (ICA) to detect Satsuma dwarf virus (SDV) was developed using colloidal gold conjugates of anti-SDV monoclonal antibodies. Of six homogenization buffers tested, 0.1 M citrate buffer (pH 7.0) gave the best results for the ICA. In the ICA, addition of 0.1% thioglycolic acid in the homogenization buffers that have been widely used in enzyme-linked immunosorbent assays (ELISA) was deleterious to the reaction because of undesirable coagulation of the colloidal gold. ICA using the anti-SDV monoclonal antibodies was 8 times and 16 times more sensitive than double antibody sandwich-ELISA and ICA using the anti-SDV polyclonal antibody, respectively. The analysis is complete in only 15 min. Furthermore, ICA using the anti-SDV monoclonal antibodies could also detect SDV-related viruses.     

Seasonal abundance and spatial distribution of the predator<Emphasis Type="Italic">Macrolophus costalis</Emphasis> and its prey<Emphasis Type="Italic">Myzus persicae</Emphasis> on tobacco

Field studies were conducted to assess the population and the spatial dynamics of the predatory bugMacrolophus costalis Fieber (Hemiptera: Miridae) and of its prey, the aphidMyzus persicae (Sulzer) (Hemiptera: Aphidoidea), on tobacco. From an untreated tobacco field in Tithorea (central Greece), tobacco leaves were collected from the upper and the lower half of the plants from June until September, in 1999 and 2000. The numbers ofM. costalis andM. persicae individuals per leaf were counted. Most aphids were observed during July and August (early and mid season), with densities dropping markedly in September. In contrast,M. costalis population densities increased late in the season (September). Significantly higher numbers of aphids were found on the upper half of the plants than on the lower half. In contrast, significantly moreM. costalis individuals were observed on the lower half. Iwao’s Regression Analysis was used in order to characterize the spatial pattern of the two species. According to this model, in both sampling seasons, aphids andM. costalis nymphs displayed an aggregated spatial pattern, whileM. costalis adults were found to be randomly distributed among sampling units. Although moreM. costalis individuals were recorded on leaves with relatively high aphid densities, this species did not react numerically to changes in prey density. In addition, a significant number of bugs were found on leaves with low aphid densities or no aphids at all. http://www.phytoparasitica.org posting Dec. 18, 2002.     

多菌灵等5种杀菌剂对烟草立枯病菌的生物活性

采用生物测定方法分析了烟草立枯病菌Rhizoctonia solani在菌丝生长、菌核形成与萌发阶段对5种杀菌剂(多菌灵、代森锰锌、菌核净、异菌脲及苯醚甲环唑)的敏感性,同时通过离体叶片法考察了5种杀菌剂对烟草立枯病的保护和治疗作用。结果表明:供试5种药剂对立枯病菌菌丝生长、菌核形成与萌发均表现出了不同程度的抑制活性,同时对烟草立枯病具有一定的保护和治疗作用。其中,对菌丝生长以及菌核形成与萌发抑制活性最强的均是多菌灵,其抑制菌丝生长的EC50平均值为(0.06 ±0.01) mg/L,0.25和20 mg/L下对菌核形成和萌发的抑制率均为100%;其次为异菌脲[EC50值为(0.35 ±0.15) mg/L,2和100 mg/L下对菌核形成和萌发的抑制率分别为(54.79 ±12.58)%和100%]、苯醚甲环唑[EC50值为(0.55 ±0.53) mg/L,5和200 mg/L下对菌核形成和萌发的抑制率分别为(97.97 ±2.64)%和100%]及菌核净[EC50值为(1.31 ±0.14) mg/L,10和100 mg/L下对菌核形成和萌发的抑制率均为100%];最弱的均是代森锰锌,其抑制菌丝生长的EC50平均值为(6 ±0.20) mg/L,10和200 mg/L下对菌核形成和萌发的抑制率分别为(43.58 ±31.87)%和0。离体试验表明:对烟草立枯病保护作用最强的是多菌灵、异菌脲和菌核净,50 mg/L时防效均 > 99%,其次为代森锰锌(50 mg/L防效为75.83%),最弱的是苯醚甲环唑(50 mg/L防效为39.29%);治疗作用最强的也是多菌灵,50 mg/L防效为93.46%,其次为菌核净、异菌脲和苯醚甲环唑(200 mg/L时防效均 > 84%),最弱的为代森锰锌(800 mg/L时防效为76.40%)。研究结果可为烟草立枯病化学防治药剂筛选提供参考和依据。     

Occurrence and incidence of viruses infecting green beans in south-eastern Spain

A study was conducted to determine the identity and prevalence of viruses in 455 greenhouses in the main Spanish green bean growing area. Directed surveys were conducted in 422 crops from 2000–2004 to collect samples from diseased plants displaying symptoms that could be attributed to viruses. The samples were analysed to detect any virus by means of dsRNA extraction, mechanical inoculation to test plants, as well as ELISA and/or RT-PCR tests to detect potyviruses, geminiviruses and viruses previously known to infect beans in Spain. Random surveys were conducted in the years 2002 and 2005 (in 21 and 12 greenhouses, respectively) to study the actual incidence of known viruses in the area. Symptoms were recorded in 23,108 plants from which 664 plants were collected and analysed by ELISA or RT-PCR. The results of the directed surveys showed that all the analyzed crops carried the cryptic virus Phaseolus vulgaris endornavirus (PVuV), whereas phytopathogenic viruses appeared in smaller percentages of the crops: Tomato yellow leaf curl virus (TYLCV) 20.4%, Southern bean mosaic virus (SBMV) 9.0%, Tomato spotted wilt virus (TSWV) 4.0%, and the new species Bean yellow disorder virus (BnYDV) that broke out in 2004 with occurrence values higher than 34.3% that year. From 2000–2004 an important decrease in TYLCV was observed, along with a slight increase in SBMV and a consistently low occurrence of TSWV. The results of the random surveys confirmed the increased occurrence of virus detected during the directed surveys, and furthermore demonstrated the percentage of incidence for each virus.     

Comparison of the effects of salicylic acid and ethephon with virus-induced hypersensitivity and acquired resistance in tobacco

The induction of a hypersensitive reaction in Samsun NN tobacco by tobacco mosaic virus (TMV) at 20°C leads to the development of both localized and systemic acquired resistance, and is associated with the appearance of pathogenesis-related proteins (PR's) and large increases in peroxidase activity and ethylene production. Salicylic acid (SA) induced a similar resistance in treated plant parts and occasionally also in untreated upper leaves of plants of which three lower leaves had been injected. SA also induced the same four PR's, but these were confined to the treated leaves. Thus, the connection between the presence of PR's and the reduction of TMV multiplication and spread may not be direct.In contrast to TMV, SA did not stimulate ethylene production and hardly increased peroxidase activity. Induction of acquired resistance and PR's by SA developed equally well at 20°C and at 32°C. However, pricking leaves with needles moistened with the ethylene-releasing compound ethephon mimicked TMV infection in inducing acquired resistance and PR's in both treated and untreated leaves at 20°C, but not at 32°C. Ethephon increased peroxidase activity at both temperatures, but only at 20°C dit it induce changes in both the anodic and the cathodic isoenzymes that were similar to those induced as a result of TMV infection. SA induced PR's and reduced TMV multiplication in Samsun tobacco, and inhibited virus spread in Samsun NN at 32°C.These observation indicate that neither the induction of PR's, nor the development of acquired resistance is temperature-sensitive. On the other hand, the effects of ethephon are temperature-sensitive in the same way as the hypersensitive response to TMV. It can thus be hypothesized that ethylene, produced naturally during the hypersensitive reaction of tobacco to TMV, leads to the temperature-sensitive synthesis or release of a presumably benzoic acid-type compound that functions as the natural inducer of PR's and acquired resistance. Although vanillic acid has been shown to accumulate in hypersensitively reacting tobacco leaves, it produced none of the effects of SA, and thus cannot be the natural inducer.Samenvatting Inductie van een overgevoeligheidsreactie in Samsun NN-tabak door tabaksmozaïekvirus (TMV) bij 20°C leidt tot de ontwikkeling van een verworven resistentie die zowel lokaal als systemisch werkzaam is, en gaat samen met het verschijnen van pathogenesis-related proteins (PR's) en sterke toename in de activitieit van peroxidase en de produktie van ethyleen. Salicylzuur (SA) induceerde een vergelijkbare resistentie in behandelde plantedelen en af en toe ook in niet behandelde bovenbladeren van planten waarvan drie onderbladeren waren ingespoten. SA induceerde ook dezelfde vier PR's, maar deze waren beperkt tot de behandelde bladeren. Er bestaat dus geen directe samenhang tussen de aanwezigheid van PR's en de remming van de vermeerdering en uitbreiding van TMV in de plant.In tegenstelling tot TMV stimuleerde SA de ethyleenproduktie niet en verhoogde het de peroxidaseactiviteit nauwelijks. Inductie van verworven resistentie en PR's door SA trad even goed op bij 32°C als bij 20°C. Net als infectie met TMV leidde aanprikken van bladeren met naalden die gedoopt waren in een oplossing van ethefon — waaruit in het blad ethyleen vrijkomt — echter tot inductie van verworven resistentie en PR's in zowel behandelde als onbehandelde bladeren bij 20°C, maar niet bij 32°C. Ethefon verhoogde de peroxidaseactiviteit bij beide temperaturen, maar alleen bij 20°C induceerde het veranderingen in zowel de anodische als de kathodische isoënzymen die vergelijkbaar waren met die welke geïnduceerd werden als gevolg van infectie met TMV. SA induceerde PR's en verminderde de vermenigvuldiging van TMV in Samsun tabak, en remde de uitbreiding van het virus in Samsun NN bij 32°C.Deze waarnemingen tonen dat noch de inductie van PR's, noch de ontwikkeling van verworven resistentie een temperatuurgevoelig proces is. Daarentegen zijn de effecten van ethefon op dezelfde wijze temperatuurgevoelig als de overgevoeligheidsreactie op TMV. Men kan daarom veronderstellen dat ethyleen, dat op natuurlijke wijze geproduceerd wordt tijdens de overgevoeligheidsreactie van tabak op TMV, aanleiding geeft tot een temperatuurgevoelig proces, namelijk de synthese of het vrijkomen van een verbinding, vermoedelijk een benzoëzuurderivaat, dat fungeert als de natuurlijke inductor van PR's en verworven resistentie. Hoewel is aangetoond dat vanillinezuur zich ophoopt in overgevoelig reagerende tabaksbladeren, veroorzaakte deze verbinding geen enkel van de effecten van SA. Vanillinezuur kan dus niet de natuurlijke inductor zijn.