Electron microscopical investigations of mosaic diseased wheat plants found in Italy

Summary For some years past at several places in the province of Brescia a mosaic disease of wheat has been detected which probably must be attributede to a virus (Grancini, 1959). Presumably the same disease has recently been constatated in other provinces of North Italy.Electron microscopical study of diseased wheat leaves showed that the preparations which were obtained with the dipping method ofBrandes (1957) all contained rod-shaped virus particles with a length of 300 m. These particles were absent from healthy plants. Whether or not this virus is directly responsible for the mosaic disease could not be established, since we did not succeed in transmitting the virus artificially.Samenvatting Sedert einige jaren wordt op verschillende plaatsen in de provincie Brescia een mozaïekziekte bij de tarwe gevonden, waarvna de oorzaak waarschijnlijk aan een virus moet worden toegeschreven (Grancini, 1959).Bij elektronemicroscopisch onderzoek van zieke bladeren is gevonden, dat de preparaten, welke met de doop-methode vanBrandes (1957) verkregen waren, alle staafvormige virusdeeltjes ter lengte van 300 m bevatten (fig. 1). In preparaten van gezond bladmateriaal kwamen deze deeltjes niet voor. Of dit virus direct aansprakelijk is voor de mozaïekziekte kon nog niet aangetoond worden, aangezien het tot nu toe niet gelukt is het virus langs kunstmatige weg over te brengen.     

Serological relationship of some European,American, and Canadian isolates of the white clover mosaic virus

Summary By means of the exchange of antisera a qualitative relationship could be established among a Dutch, some American, a Canadian and a German isolate of the white clover mosaic virus (fig. 1). An exact comparison of two isolates, viz. the Dutch W.K.V. and the American W.C.M.V. (ofBancroft), with a number of different antisera (table 1), and in a cross-absorption test (table 2), revealed no quantitative differences between these isolates. For such quantitative tests the exchange of virus isolates is inevitable.Samenvatting Door middel van de uitwisseling van antisera kon een kwalitatieve verwantschap worden aangetoond tussen een Nederlandse, enige Amerikaanse, een Canadese en een Duitse isolatie van het witte-klavermozaïekvirus. Een nauwkeurige vergelijking van twee isolaties, te weten het Nederlandse W.K.V. en het Amerikaanse W.C.M.V. (vanBancroft), met behulp van een aantal verschillende antisera (tabel 1) en in een proef, waarbij de antisera met wederzijdse virussen werden verzadigd (tabel 2), bracht geen kwantitatieve verschillen (stam-verschillen) aan het licht. Voor dergelijke kwantitatieve proeven is de uitwisseling van virusisolaties onvermijdelijk.Journal Paper No. 1559 of the Purdue University Agricultural Experiment Station.     

Serological variability among European isolates of radish mosaic virus

Serological variability of radish mosaic virus (RaMV) isolates from white mustard, winter turnip rape, Camelina sativa and Chinese cabbage, collected in the Czech Republic and Russia, was studied using antisera against the Czech isolate RaMV1. In contrast to previous studies, reasonable serological differences were found between isolates from different locations and hosts, and even between neighbouring mustard plants in the field. Serological variability of European isolates was confirmed in extended experiments involving an Italian isolate, the American Type Strain (ATS) and its homologous antiserum. The results indicate that, in Europe, RaMV isolates may occur which differ both serologically and in host plant response, but no typical strains can be defined with the methods and isolates employed so far. Camelina sativa is reported as a new host of the virus.     

Light and electron microscopy of red clover vein mosaic virus in pea (Pisum sativum)

Epidermal strips of pea infected with the RK31 isolate of the red clover vein mosaic virus were found to contain many diagnostic crystalline inclusions, recognizable two, but more clearly three weeks after inoculation. They could be easily detected by light microscopy, even at low magnification, when stained with phloxine and methylene blue, but more rapidly with a phase-contrast microscope in unstained preparations in water. They were always found after the onset of external symptoms. With the pea streak strain (P42) they were usually absent. In ultrathin sections made 22 days after inoculation or later, virus distribution and accumulation were comparable to those of other members of the carlavirus group. Sometimes extensive irregular bundles were observed. These were distinct from the crystals seen by light microscopy. Sections of the three-dimensional crystals studied in the electron microscope showed a very regular internal striation with a periodicity of about 11 nm, or were sometimes composed of spherical particles in looser array. When crystals were isolated intact and stained negatively, the majority of the material appeared to consist of spherical or polyhedral particles 11–12 nm in diameter.     

大麦条纹花叶病毒中国株三基因盒(TGB)序列对比分析

 The tripe gene block (TGB)genes of Barley stripe mosaic virus China strain (BSMV-CH)were amplified from cDNA of BSMV-CH RNAβ (GenBank accession No:AY789694)by PCR with special primer pairs, and cloned into pMD18-T vector for sequencing. Analysis of the sequences showed that the full length of BSMV-CH TGB1, TGB2 and TGB3 were 1 539, 396 and 468 bp, with deduced 512, 131 and 155 amino acids, respectively. BSMV-CH TGB1 shared 94.1%-95.4% nucleotide identities and 91.0%-94.5% amino acid identities with that of other BSMV strains, BSMV-CH TGB2 shared 96.5%-97.2% nucleotide identities and 98.5%-99.2% amino acid identities with that of other BSMV strains, and BSMV-CH TGB3 shared 95.7%-96.6% nucleotide identities and 94.2%-96.8% amino acid identities with that of other BSMV strains. Phylogenetic tree based on the amino acid of Hordeiviruses TGB genes showed that BSMV-CH was relatively closed to CV17 in genetic relationship. Therefore, BSMV-CH was deduced to be a recombined strain of CV17 and CV42.     

大麦条纹花叶病毒荧光定量PCR检测方法的建立及应用

大麦条纹花叶病毒Barley stripe mosaic virus（BSMV）是我国进境植物检疫性有害生物。为提高大麦条纹花叶病毒检测的灵敏度和特异性,缩短检测周期,根据该病毒不同分离株外壳蛋白（coat protein, CP）基因的保守序列设计特异性的引物和探针,建立了BSMV的实时荧光RT-PCR检测方法。结果表明,本检测方法特异性强,对南方菜豆花叶病毒Southern bean mosaic virus（SBMV）、小麦线条花叶病毒Wheat streak mosaic virus（WSMV）、玉米褪绿斑驳病毒Maize chlorotic mottle virus（MCMV）、烟草环斑病毒Tobacco ringspot virus（TRSV）、菜豆荚斑驳病毒Bean pod mottle virus（BPMV）和番茄环斑病毒Tomato ringspot virus（ToRSV）6种病毒均无交叉扩增;且检测方法灵敏度高,对RNA模板的最低检测限达到5×10-4 ng/μL,与双抗体夹心酶联免疫吸附法（DAS-ELISA）和普通RT-PCR相比,本方法的灵敏度分别提高了10倍和100倍。此外,我们利用该方法对12批进口大麦进行检测,发现6批大麦中检出大麦条纹花叶病毒,占比约50%。总之,本研究建立的荧光定量PCR方法具有灵敏度高、特异性强等优点,适合于BSMV的快速检测。     

Progress towards the integrated control of sugar cane pests

Sugar cane is an important tropical and sub-tropical crop that occupies large areas of land around sugar factories capable of processing 5000 to 15 000 tonnes of cane per day. These areas are usually kept continuously under the same crop, about one-tenth to one-third of the land being ploughed and replanted with sugar cane every year. The major pests of sugar cane are rats, stalk borers, white grubs and froghoppers. Other important pests include root-feeding nematodes, top borers, scale insects, planthoppers and soldier fly. Chemical control is practised to some extent against most of these pests, although biological control is the most common method of combating stalk-borers and has had other notable successes, particularly in Hawaii and Mauritius. Rats are controlled with baits incorporating poisons such as anticoagulants, endrin or zinc phosphide. Aerial applications of insecticide sprays, dusts or granules are used against some stalk-borers (azinphos-methyl), froghoppers (omethoate, propoxur, vamidothion, methidathion, monocrotophos, malathion, carbaryl, gamma-HCH) and plant hoppers (malathion, fenitrothion, endrin). White grubs, soldier fly (crude HCH, dieldrin) and nematodes [1,2-dibromoethane (EDB), 1,2-dibromo-3-chloropropane (DBCP), aldicarb] are usually controlled with soil applications of the appropriate chemicals. The use of chemical control is restricted in some areas because it is too costly and in other areas because of its possibly adverse effects upon the natural or biological control of other pests. Historically, pest control in sugar cane has tended to develop towards a simple integrated system, making use of cultural methods in conjunction with chemicals or exotic parasites and predators. Increasingly, allowance has been made for the contribution of indigenous natural enemies to the prevention of crop losses by a more careful, selective use of insecticides.     

Serological and biological variation between and within subgroup I and II strains of cucumber mosaic virus

Fourteen strains of cucumber mosaic virus (CMV) from Australia have been characterized by their host range and symptomatology. They were classified as subgroup I or II strains by a dot-blot molecular hybridization assay between their total viral RNAs and selected cDNAs. The strains F_NY and L_Ny, both from the USA, were used as the subgroup I- and subgroup II-type strains, respectively. A range of serological tests was used to compare these isolates. Gel immunodiffusion tests, with standard antigens homologous to the antisera prepared against glutaraldehyde-fixed virus of 11 strains, showed that they could be divided into three serogroups on the basis of spur formation in heterologous reactions. Two of the serogroups included either subgroup I or subgroup II isolates, whereas the third serogroup consisted of only one strain (Y_WA) which was homologous to all the strains tested. Use of heterologous standard antigens in this test failed to show further subgrouping of the antigens. Double-antibody sandwich (DAS) ELISA using polyclonal antibodies to distinct virus strains also placed the 14 strains in the same three serogroups. When eight different monoclonal antibodies (MAbs) were used in indirect ELISA, one of them distinguished subgroup-I strains and another distinguished subgroup-II strains; the Y_WA strain fell into subgroup II. Other MAbs showed narrower or broader specificity. Thus both molecular hybridization with total RNA and specific MAbs may be useful for separating isolates of CMV into subgroups I and II. Spur formation using heterologous standard antigens to the antisera, as well as being more difficult to interpret, was not a reliable criterion for classification.     

Serological differences between red currant spoon leaf virus,virus isolates from Eckelrade-diseased cherry trees and the Scottish raspberry ringspot virus

Antisera were made to red currant spoon leaf virus (SLV), an isolate of the Scottish raspberry ringspot virus (RRV), and two virus isolates from Eckelrade-diseased cherry trees (EV). Different virus isolates, including one from Belgium, were tested against these antisera. The results indicate that we are dealing with a group of virus isolates with different antigenic properties. SLV is very closely related to RRV, being undoubtedly a strain of this virus. Dutch EV isolates differ from SLV and RRV and from each other, the Belgian isolate being closely related to one of the Dutch EV isolates. The serological differences found do not correspond with the geographical distances between the localities where the viruses were collected.Samenvatting Antisera werden bereid tegen het lepelbladvirus van rode bes (SLV), een isolatie van het Schotse raspberry ringspot-virus (RRV) en twee virusisolaties uit kersebomen met Eckelraderziekte (EV). Met behulp van deze antisera werden verschillende virusisolaties getoetst, waaronder één uit België van kers met Eckelraderziekte. De resultaten zijn vermeld in de tabellen 1 en 2. Ze duiden erop, dat we te maken hebben met een groep van virusisolaties met verschillende antigene eigenschappen. SLV is zeer nauw verwant aan RRV en is ongetwijfeld een stam van dit virus. Nederlandse EV-isolaties verschillen zowel van SLV en RRV, als van elkaar. De Belgische isolatie is nauw verwant aan één van de Nederlandse EV-isolaties. De grootte van de gevonden verschillen correspondeert niet met de geografische afstand tussen de plaatsen waar de virusisolaties werden verzameld.     

灰飞虱不同虫量与水稻条纹叶枯病发病的相关性研究

通过田间水稻1代灰飞虱成虫高峰期罩笼接虫传毒试验,结果表明,带毒灰飞虱属持久性传毒型介体昆虫,并随虫量的增加,水稻条纹叶枯病发病呈上升趋势,以400～600头/m2时发病较为明显。浸种试验显示,用25%咪鲜胺EC(使百克) 10%二硫氰甲烷EC(浸种灵) 12.5%吡虫啉SL浸稻种有控虫防病效果。     

A Comparison of Ultrastructural Changes of Barley Cells Infected with Mild and Aggressive Isolates of Barley stripe mosaic virus

Journal of Plant Diseases and Protection - Electron microscopy studies were carried out to investigate the association of visible disease symptoms with ultrastructural changes in barley plants...     

Economic impact of Turnip mosaic virus, Cauliflower mosaic virus and Beet mosaic virus in three Kenyan vegetables

Screenhouse experiments conducted in Kenya showed that inoculation of cabbage seedlings with Turnip mosaic virus (TuMV), either alone, or in combination with Cauliflower mosaic virus (CaMV), reduced the number and weight of marketable harvested heads. When viruses were inoculated simultaneously, 25% of cabbage heads were non-marketable, representing 20-fold loss compared with control. By contrast, inoculation with CaMV alone had insignificant effects on cabbage yield. This suggests that TuMV is the more detrimental of these pathogens, and its management should be a priority. Early exposure to TuMV produced cabbages that were 50% lighter than non-infected plants, but later infection was less damaging suggesting that controlling virus infection at the seedling stage is more important. TuMV was far less damaging to kale than it was to cabbage; although high proportions of TuMV-inoculated kale plants showed symptoms (>90%), the marketability and quality of leaves were not significantly reduced, and no clear relationship existed between timing of infection and subsequent crop losses. Early inoculation of Swiss chard with Beet mosaic virus (BtMV) significantly impaired leaf quality (∼50% reduction in marketable leaf production), but the impact of disease was greatest in plants that had been inoculated at maturity, where average leaf losses were two and a half times those recorded in virus-free plants. Disease-management of BtMV in Swiss chard is important, therefore, not only at the seedling stage, but particularly when plants are transplanted from nursery to field.     

西瓜感染黄瓜绿斑驳花叶病毒后糖的变化与倒瓤关系的研究

 By high performance liquid chromatography (HPLC), the contents of glucose, fructose and sucrose, and the sweetness were analyzed in watermelon inoculated with Cucumber green mottle mosaic virus (CGMMV).The relationship between sugar change and blood-flesh of watermelon was determined. The results indicated that the content of glucose was apparently increased compared to that of the control before maturity (within 28 days after pollination), but reduced to 24.8% of the control after maturity (at 35 days after pollination). The content of fructose was higher than that of the control within 14 days after pollination, and then decreased with a significant difference. In all cases, the content of sucrose increased with the growth of watermelon. However, compared with the control, the sucrose content of watermelon inoculated with CGMMV was lower. The ratios of glucose, fructose and sucrose in the total sugar were abnormal. Coincidence with the changes of the total sugar, the fruit sweetness before maturity was higher than that of the control, whereas decreased sharply after maturity (lower than that of the control). The inner pulp of the mature fruit appeared to be water-soaked and dirty red with no edibility. In a word, after inoculation with CGMMV, the changes of sugars and sweetness affected the watermelon quality.     

Evaluation of Maize Inbreds for Maize stripe virus and Maize mosaic virus Resistance: Disease Progress in Relation to Time and the Cumulative Number of Planthoppers

ABSTRACT Five tropical maize lines were tested and compared with the susceptible control line B73 for resistance to Maize stripe virus (MStV) and Maize mosaic virus (MMV), both propagatively transmitted by the planthopper Peregrinus maidis (Homoptera: Delphacidae). Resistance to each virus was evaluated separately by artificial inoculations with planthoppers viruliferous for either one virus or the other. Disease incidence and symptom severity progression were quantified in relation to time and the cumulative number of planthoppers. Line Hi40 was found to be susceptible to MStV and highly resistant to MMV. Generally, no MMV symptoms developed on Hi40, even under intense inoculation pressure by a large number of viruliferous planthoppers. Line Rev81 showed a partial but strong resistance to MStV, which mainly reduced disease incidence. Nevertheless, this resistance to MStV was the highest ever reported and held up, even when challenged by large numbers of planthoppers. The percentage of infected plants in line Rev81 never exceeded 30 to 40% in our experiments. Moderate levels of resistance to MStV, and to a lesser extent MMV, were found in lines 37-2, A211, and Mp705. However, resistance in these lines was completely overcome using a large number of insects transmitting either of the two viruses. These results suggest that different types of resistance to MMV and MStV are available in maize lines from Caribbean and Mascarene germ plasm. The expression of virus-specific resistance identified in Hi40 and Rev81 lines was not affected by intense inoculation pressure. In contrast, the moderate resistance in 37-2, A211, and Mp705 was partially effective against both viruses but not at high inoculation pressure. These different types of resistance, when present in the same genotype, could provide protection against both viruses.     

硝磺草酮在甘蔗和土壤中的残留行为

建立了硝磺草酮在甘蔗和土壤中的残留分析方法,并在广东和广西分别进行了10%硝磺草酮悬浮剂在甘蔗上残留的田间试验,研究了硝磺草酮在甘蔗和土壤中的消解动态和最终残留量。样品用乙腈提取,盐酸调节pH至3~4后二氯甲烷萃取,采用高效液相色谱配二极管阵列紫外检测器(HPLC-PDA)检测。结果表明:在0.01、0.1和1 mg/kg 3个添加水平下,硝磺草酮平均添加回收率为82%~84%,相对标准偏差(RSD)为1.9%~3.4%,检出限(LOD)为0.005 mg/kg,定量限(LOQ)为0.01 mg/kg。田间试验结果表明:施用10%硝磺草酮悬浮剂后,在甘蔗上的残留量呈现先降低后升高又下降的趋势;在土壤中的半衰期为12.3~14.7 d,属于易降解农药。最终残留量测定结果显示:收获期甘蔗中硝磺草酮的残留量均0.01 mg/kg。     

地高辛标记的cDNA探针检测烟草花叶病毒、黄瓜花叶病毒及马铃薯Y病毒

 根据已发表的烟草花叶病毒(Tobacco mosaic virus,TMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)和马铃薯Y病毒(Potato virus Y,PVY)的外壳蛋白基因序列,设计特异引物,分别以提取的TMV、CMV和PVY侵染的病叶总RNA为模板,反转录PCR进行体外扩增,分别得到长度为0.44、0.77、0.80 kb的目的片段,并克隆到pGEM-T easy质粒载体上,以构建的重组质粒为模板,用PCR方法合成了相应的地高辛标记的双链DNA探针。以合成的探针通过斑点杂交技术检测烟草病叶总RNA和烟草病叶汁液。TMV、CMV和PVY的3种地高辛探针检测各自感染的烟草病叶总RNA的稀释低限分别为1:1000、1:10000、1:320,检测各自侵染烟草病汁液的最大稀释倍数分别为1:100、1:100、1:10,而每种探针与健康烟草和其它2种病毒的反应均为阴性。     

噻唑磷在甘蔗和土壤中的残留消解动态及风险评估

建立了噻唑磷在甘蔗和土壤中的残留分析方法,并在广东和广西2地进行了10%噻唑磷颗粒剂在甘蔗上残留的田间试验,研究了噻唑磷在甘蔗和土壤中的消解动态和最终残留量,并对甘蔗中噻唑磷可能产生的膳食摄入风险进行了评估。甘蔗样本用乙腈提取,氨基固相萃取小柱净化,采用气相色谱-火焰光度检测器 (GC-FPD) 检测。结果表明:在0.01、0.1和1 mg/kg添加水平下,噻唑磷在甘蔗中的平均回收率为83%～84%,相对标准偏差 (RSD) 为2.6%～3.4%;在土壤中的平均回收率为84%～86%,RSD为4.1%～6.7%。噻唑磷检出限 (LOD) 和定量限 (LOQ) 均为0.01 mg/kg。田间试验结果表明:10%噻唑磷颗粒剂在广东和广西甘蔗中的消解不符合一级反应动力学方程,没有显著的消解规律,呈现的特点是浓度由小到大再变小的趋势,施药后14～21 d甘蔗中噻唑磷的残留量达到最大值0.027 mg/kg;噻唑磷在土壤中消解符合一级反应动力学方程,半衰期为8.6～9.6 d,属易降解农药;噻唑磷在甘蔗和土壤中的最终残留量均小于0.01 mg/kg。膳食摄入风险评估结果表明:甘蔗中噻唑磷对人群的急性膳食摄入风险值为225%,急性膳食摄入风险较大,还需进一步结合噻唑磷在人体内的代谢行为等数据进行综合评估;慢性膳食摄入风险值为57%,表明按照推荐剂量施用10%噻唑磷颗粒剂,慢性膳食摄入风险较低,对消费者健康是安全的。