A method to test wheat leaves for their reactions to inoculation with Septoria species

The construction and application is described of a polystyrol humidity box in which wheat leaves, while continuing to function as parts of living plants, can be tested for their reactions toSeptoria spp. in an atmosphere nearly saturated with water, as is required for successful infection. The method is simple, accurate ans inexpensive.Samenvatting Voor dit doel is een z.g. vochtdoos geconstrueerd (Fig. 1A). Het te toetsen blad wordt daar doorheen geleid en in de doos geïnoculeerd met een druppel conidiën-suspensie van de schimmel. Onder het blad staat wat water in de doos (Fig. 1B). Na afsluiting ontstaat in de doos de hoge luchtvochtigheid van bijna 100% die nodig is voor de infectie. Dit wordt op deze wijze eenvoudig en goedkoop gerealiseerd. De bladeren die zo worden getoetst blijven onbeschadigd functioneren aan de plant. Na enkele dagen kan de doos worden geopend en kan de symptoom-ontwikkeling worden afgewacht en gevolgd (Fig. 3A, 3B en 4). De methode leent zich voor nauwkeurig werk en vereist zeer weinig infectiemateriaal. De Technische en Fysische Dienst voor de Landbouw (TFDL), Wageningen, ontwierp en construeerde een statief voor het gebruik van de vochtdozen in serie (Fig. 2).     

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.     

水稻苗田化学除草剂—封闭1号大面积苗田药效示范试验结果

通过多点水稻苗田大面积示范试验结果表明,通化市农科所研制的水稻苗田除草剂“封闭1号”具有除草效果好(90～100％)、安全、使用方便、成本低等特点。1988～1991年,累计推广面积(苗田)达15.6万亩,创造经济效益150万元。     

Bacterial diseases of potatoes: from classical phytobacteriology to molecular pathogenicity

Various bacterial pathogens attack potato plants and tubers. These pathogens includeErwinia, Corynebacterium (Clavibacter), Psuedomonas andStreptomyces. Over the past few years there have been significant advances in the molecular biological analysis of several of these pathogens and this is now helping us to understand the major aspects of virulence mechanisms. However, to date, such information is not sufficiently useful to allow us to intervene rationally in these potato diseases, except by the standard practises of good husbandry.     

Pelargonium flower-break and pelargonium line pattern viruses in the Netherlands; purification,antiserum preparation,serological identification,and detection in pelargonium by ELISA

Two viruses, detected frequently in the Netherlands in pelargonium, were identified by serology and test plant reactions. Antisera were prepared and an ELISA procedure was developed to detect the viruses in pelargonium.One of the viruses, PFBV-N, proved to be pelargonium flower-break virus. With the antiserum to PFBV-N, it could be detected reliably throughout the year inPelargonium zonale Springtime Irene.The other virus, PLPV-N, was serologically closely related to pelargonium line pattern virus (PLPV) and to pelargonium ring pattern virus (PRPV), as were an old virus isolate from Saturnus, collected in the Netherlands in 1971 (L128), and PLPV isolates from Yugoslavia (PLPV-Y) and Denmark (PLPV-D). There were only minor differences in host-plant reactions between the virus isolates. Based on these tests, PLPV and PRPV are considered as isolates of the same virus, for which, for practical reasons, the name pelargonium line pattern virus is proposed.PLPV could be reliably detected by ELISA inP. zonale Springtime Irene and Amanda throughout the year with only a few exceptions. InPelargonium peltatum Tavira, however, reslts were erratic due to uneven distribution of virus in the plant. Best results were obtained when petioles of fully expanded leaves were tested.     

Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA)

Squash mosaic virus (SqMV, comovirus) is seed-transmitted in severalCucurbitaceae. Therefore, the use of virus-free seed is important to prevent establishment of this virus in the Netherlands and to avoid spread to other countries.This study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of SqMV in melon seeds. An antiserum was produced to a serotype 1 isolate from melon. Two ELISA variants were investigated viz. an ELISA variant with simultaneous incubation of sample and enzyme conjugate (ELISA 1) and an ELISA variant with successive incubation of sample and enzyme conjugate (ELISA 2). The sensitivity of ELISA was tested by mixing fluor of ground infected and non-infected seeds in different proportions. SqMV was detected by both ELISA variants at dilutions of 1 160 (1 part of infected flour mixed with 159 parts of non-infected flour) or higher after a substrate incubation period of 4 h. However, ELISA 1 gave relatively higher absorbance values than ELISA 2 for nearly all dilutions. Since ELISA 1 is also faster than ELISA 2, ELISA 1 is advised for routine testing. In these test, using subsamples of 100 melon seeds SqMV is detected reliably. ELISA 1 is now used in the Netherlands for routine-indexing of melon seed lots for SqMV.Samenvatting Het pompoenemozaïekvirus gaat over met het zaad van verscheideneCucurbitaceae. Het gebruik van virusvrij zaad is belangrijk om te voorkomen dat het virus zijn intrede doet in Nederland en zich naar andere landen verspreidt.Een antiserum werd geproduceerd tegen een serotype 1 isolaat van meloen. Met behulp van dit antiserum werd een ELISA ontwikkeld om pompoenemozaïekvirus in zaden van meloen aan te tonen. Twee varianten van ELISA werden vergeleken, namelijk een variant waarbij monster en enzymconjugaat gelijktijdig geïncubeerd werden (ELISA 1) en een variant waarbij monster en enzymconjugaat na elkaar geïncubeerd werden (ELISA 2). De gevoeligheid van de ELISA varianten werd uitgetoetst door meel van zieke zaden in verschillende verhoudingen te mengen met meel van gezonde zaden. Het pompoenemozaïekvirus werd met beide ELISA varianten aangetoond in verdunningen van 1 160 (1 deel meel van zieke zaden gemengd met 159 delen meel van gezonde zaden) of hoger na 4 uur incubatie met substraat. ELISA 1 gaf doorgaans hogere extinctiewaarden dan ELISA 2 voor bijna alle verdunningen. Omdat ELISA 1 ook nog sneller is dan ELISA 2, wordt ELISA 1 aanbevolen voor routinematig gebruik. Wanneer voor routinematig gebruik 100 meloenezaden per submonster getoetst worden, kan het pompoenemozaïek virus betrouwbaar worden aangetoond. In Nederland worden momenteel per zaadpartij 20 submonsters van 100 zaden getoetst.     

A seroepidemiological study on bluetongue virus in dairy cattle in the central valley of California

A seroepidemiological study on bluetongue virus (BTV) infection in California dairy cattle was conducted to estimate the prevalence and distribution by age and season of BTV group-reactive antibodies and to look for possible associations between the presence of antibodies and cattle age or breed and farm. Between December 1985 and March 1987, a sample of cattle was tested at approximately two-month intervals for BTV group-reactive antibodies using an enzyme-linked immunosorbent assay (ELISA). Data taken during the month of December 1986 were used to evaluate possible associations between a positive antibody test and certain intrinsic (age, breed) and extrinsic (farm) factors.Univariate and multivariate statistical analyses using the -square test for associations and multiple logistic regression, respectively, were carried out for possible associations between positive antibody tests to BTV and each factor of interest. The strengths of the associations were determined using estimates of the odds ratio.Of the 3774 serum samples tested, 238 (6.3%) were from calves, 1045 (27.6%) were from heifers and 2492 (66.0%) were from cows. Seroprevalence varied from nil in calves on two occasions to over 90% on several occasions in cows. Cows consistently had higher prevalence rates than heifers or calves across all test dates (p<0.05). The seroprevalence of BTV group-reactive antibodies also showed a seasonal fluctuation, with the highest rates occurring during the warmer months of the year. These highest prevalence rates coincided with heavy activity of the known vector of BTV, Culicoides spp. Breed and farm effects were not statistically significant (p>0.05). With the exception of one farm, all cattle were of the Holstein breed, which reduced confidence in assessing any breed effect in this study. Relative estimates of the sensitivity and specificity of BTV ELISA were 87% and 100% respectively, compared to the standard agar gel immunodiffusion (AGID) test.The observations support previous findings of seasonal distribution of BTV antibodies and suggest an age relationship, whereby older cattle are more likely to be positive to BTV group-reactive antibodies than younger cattle.     

ONCHOCERCA GUTTUROSA IN Danish cattle. Prevalence, geographic distribution and host-vector relationships.

1. Onchocerca gutturosa is reported in Danish cattle for the first time. Microfilariae were found in 38 (9.4%) of 406 cows that were 2 years or older.
2. Microfilariae were not randomly distributed throughout the skin but were concentrated in the umbilical area.
3. No difference in prevalence was observed between breeds of cattle.
4. Most of the infected cows had grazed on fields close to streams that contained Simulium ornatum, the vector of O. gutturosa.
5. Differences in preferred biting sites of S. ornatum from cow to cow were correlated with the arrangement of hair.

     

自由放养下绵羊的营养参数

根据绵羊的营养参数和饲养标准,结合日增重水平,计算了不同日增重水平对应的体重月平均动态及其所对应的粗蛋白日需要量和代谢能日需要量。不同日增重水平,达到生长期望所需要的日数不同,日增重水平越高,达到生长期望总所需要的粗蛋白和代谢能越少,但月平均的日需要量越多。自由放养的东北细毛羊当年生羔羊的日增重水平仅相当于舍饲饲养日增重的100g水平,根据代谢能计算,年总需干草340kg,可以作为计算载畜量的基本参数。自由放养情况下,即使能量满足需要,粗蛋白也短缺。当年生羔羊日增重在150￣200g,9￣10月龄内体重达到46￣50kg是北方草地家畜生产的理想目标,通过补饲可以实现。     

茭白肉质茎膨大过程中的糖代谢与激素含量变化

 以单季茭白品种‘蒋墅茭’和双季茭白品种‘葑红早’为试材, 研究了秋季肉质茎膨大过程中物质运转的相关生理生化变化。结果表明, 随着肉质茎鲜质量、干质量快速增加, 非结构性碳水化合物主要成分含量总体上升, 后期出现下降, 蒋墅茭肉质茎中蔗糖含量较高; 糖分转化关键酶活性总体较高,转化酶活性以酸性转化酶较高且呈快速下降, 蒋墅茭的酸性转化酶活性高于葑红早而中性转化酶活性较低;蔗糖合成酶的分解方向活性显著高于合成方向, 且后期维持较高水平, 葑红早的双向活性均高于蒋墅茭;Z + ZR总体含量最高、下降最明显, GA₄ 其次, IAA也呈缓慢下降趋势, GA₃、ABA含量极低, 且各激素均以葑红早高于蒋墅茭。茭白肉质茎干、鲜质量处于快速增长阶段的同时, 而内部生理生化快速下降, 说明茭白肉质茎膨大过程具有特殊性, 并且品种间存在差异。