Elimination of Sugarcane yellow leaf virus from infected sugarcane plants by meristem tip culture visualized by tissue blot immunoassay

Sugarcane yellow leaf virus (SCYLV), a member of the Luteoviridae , is implicated in the sugarcane disease known as yellow leaf syndrome (YLS), which is characterized by yellowing of the leaf midrib followed by leaf necrosis and possible growth suppression. YLS is distributed worldwide and susceptible cultivars are commonly infected with SCYLV. However, not all cultivars infected with SCYLV show symptoms of YLS and some cultivars that show symptoms do so sporadically. Since it is difficult to obtain virus-free plants of susceptible cultivars, it has not been possible to study the factors involved in SCYLV infection nor the effects of infection on plant growth and yield. A tissue blot immunoassay was used to visualize in vivo presence of the virus so that virus-infected and virus-free plants could be distinguished. Meristem tip cultures were used to produce virus-free plantings of six SCYLV-susceptible sugarcane cultivars. Nearly all of the regenerated sugarcane lines remained virus-free over a period of up to 4 years, whether grown in isolated fields or in the glasshouse. Experimental re-infection of the virus-free plants by viruliferous aphids demonstrated that meristem tip culture did not affect susceptibility of sugarcane to SCYLV. Improved diagnosis and production of virus-free plants of SCYLV-susceptible cultivars will facilitate research to quantify the effect of the virus on yield and to analyse the processes involved in disease development.     

The host ranges,classification and identification of eight persistent aphid-transmitted viruses causing diseases in legumes

Seventy one seed-lines representing 23 species of papilionoid legumes and 17 species of nonlegumes were collected and distributed to four countries; Australia, New Zealand, the Netherlands and the U.S.A. In each of these countries plants were grown from the seeds and their susceptibility to a total of eight persistent aphid isolates transmitted viruses was assessed. The viruses were a strain of beet western yellows (BWYV) fromGlycine max in Illinois, legume yellows (LYV) in California and a virus in Michigan (MiAV) fromMedicago sativa, fromPisum sativum causing leaf roll and top yellows in New Zealand (PeLRV-NZ) and the Netherlands (BLRV), isolates of subterranean clover red leaf from New Zealand (SCRLV-NZ) and Tasmania (SCRLV-T), and subterranean clover stunt (SCSV) from Tasmania.The relationships between the eight viruses as indicated by their host reactions were assessed using computer classification techniques. SCRLV-NZ and SCRLV-T were the most similar. They had moderately wide host ranges that included some non-legumes. A second group comprized BWYV and PeLRV-NZ. These were typical of most beet western yellows virus strains in that they infectedBrassica napus, Capsella bursa-pastoris andStellaria media. MiAV and BLRV also formed a pair. They generally induced severe symptoms on the hosts which they infected and had host ranges confined to legumes except that BLRV also infectedClaytonia perfoliata andErodium spp. The relationships of LYV and SCSV were not consistent. They paired together in some classifications, but SCSV sometimes grouped with the SCRLV isolates. Both had host ranges confined to legumes, caused severe symptoms in most hosts and were often difficult to recover from affected plants. LYV had some affinities with BLRV and MiAV.The tests indicated a set of test plants which were most useful for propagating and identifying persistent aphid-transmitted viruses from legumes. Two,P. sativum cv. Onyx andTrifolium subterraneum cv. Bacchus Marsh were susceptible to all isolates. Ten others distinguished between the isolates and wereArachis hypogea, Beta vulgaris, C. bursa-pastoris, G. max cv. Shirotsurunoko,Gomphrena globosa, Lactuca sativa, Lens esculenta cv. 179307,M. sativa cv. Washoe,Phaseolus vulgaris cv. Canadian Wonder andTrifolium hybridum.Samenvatting Eenenzeventig zaadmonsters van 23 soorten van de Papilionaceae en 17 andere soorten werden verzameld en naar laboratoria in Australië, Nieuw Zeeland, Nederland en de Verenigde Staten van Amerika gezonden. In elk van deze landen werden de uit de zaden verkregen planten op hun vatbaarheid voor acht door bladluizen op persistente wijze overgebrachte virussen getoetst. Het betrof de volgende virussen: beet western yellows virus (BWYV) vanGlycine max uit Illinois, legume yellows virus (LYV) uit Californië, een virus vanMedicago sativa uit Michigan (MiAV), twee virussen die bladrol en topvergeling inPisum sativum veroorzaken, één uit Nieuw Zeeland (PeLRV-NZ) en één uit Nederland (BLRV), isolaten van het subterranean clover red leaf virus uit Nieuw Zeeland (SCRLV-NZ) en Tasmanië (SCRLV-T) en subterranean clover stunt virus (SCSV).De verwantschap tussen de acht genoemde virussen, zoals blijkend uit de reacties van de verschillende waardplanten, werd vastgesteld met behulp van een computerprogramma voor classificatie.SCRLV-NZ en SCRLV-T vertoonden de meeste verwantschap. Beide hadden een tamelijk grote waardplantenreeks, waaronder enkele soorten die niet behoorden tot de vlinderbloemigen.BMYV en PeLRV-NZ vormden een tweede groep. Zij waren typisch voor de meeste stammen van het beet western yellows virus in zoverre zij de toetsplantenBrassica napus, Capsella bursa-pastoris enStellaria media konden infecteren.MiAV en BLRV vormden eveneens een paar. Zij veroorzaakten meestal verschillende typen symptomen op de vatbare waardplanten. De waardplantenreeks was beperkt tot de Leguminosae. Alleen kon BLRV ookClaytonia perfoliata enErodium spp. infecteren.De mate van verwantschap tussen LYV en SCSV was niet eenduidig. Uit bepaalde eigenschappen zou tot onderlinge verwandtschap besloten kunnen worden, uit enkele andere echter zou meer een verwantschap tussen SCSV en SCRLV worden vermoed. De waardplantenreeks van LYV en SCSV was beperkt tot de Leguminosae; beide veroorzaakten hevige symptomen in hun waardplanten en konden moeilijk worden geïsoleerd uit geïnfecteerde planten. LYV vertoonde enige verwantschap met BLRV en MiAV.Op grond van de resultaten van de proeven kon een groep van toetsplanten worden samengesteld, die zeer bruikbaar is voor de virusvermeerdering en voor de identificatie van de genoemde acht virussen. Twee van de toetsplanten, nl.P. sativum enTrifolium subterraneum cv. Bacchus Marsh bleken vatbaar voor alle isolaten. Tien andere, nl.Arachis hypogea, Beta vulgaris, C. bursa-pastoris, G. max cv. Shirotsurunko,Gomphrena globosa, lactuca sativa, Lens esculenta cv. 179307,M. sativa cv. Washoe,Phaseolus vulgaris cv. Canadian Wonder enTrifolium hybridum zijn geschikt voor onderscheiding van alle genoemde isolaten.     

猕猴桃植株中柑橘叶斑驳病毒实时荧光定量PCR检测技术的建立及应用

 柑橘叶斑驳病毒(Citrus leaf blotch virus, CLBV)在陕西省栽培猕猴桃中发生普遍。为监测CLBV发生情况,本研究建立了CLBV的实时荧光定量PCR(Real-time fluorescent quantitative polymerase chain reaction,RT-qPCR)检测方法。该方法特异性强,可准确检测目的病毒,标准曲线斜率为-3.378,决定系数R²=0.997 9,扩增效率为97.7%,比普通RT-PCR灵敏度高100倍,可用于猕猴桃植株CLBV的批量检测或低丰度病毒样本(如猕猴桃休眠枝条)的检测。为苗木携带CLBV病毒的早期诊断、果园病毒病预测预报和防控奠定了基础。     

北京地区番茄黄化曲叶病毒病的鉴定及防治对策

番茄黄化曲叶病毒病是一种由烟粉虱传播的病毒病,给番茄生产造成严重威胁。2009年在北京郊区调查时发现部分保护地种植的番茄植株表现典型黄化曲叶症状。通过提取典型症状样品总DNA利用粉虱传双生病毒检测简并引物PA/PB,进行PCR扩增到541bp的特异条带。通过测序和核苷酸序列比对表明该序列与番茄黄化曲叶病毒序列相似性最高为99%。分子检测结果表明北京郊区部分保护地种植的番茄已被烟粉虱传播的番茄黄化曲叶病毒侵染危害。     

Biological and molecular characterization of Potato yellow blotch virus,a new species of the genus Potyvirus

A new species of the genus Potyvirus infecting potatoes, with the proposed name Potato yellow blotch virus (PYBV), was discovered in a breeding line 99m-022-026 in Scotland. The infected plants show isolated yellow blotches on the leaves. The genome of PYBV contains a large open reading frame encoding a single polyprotein of 3054 amino acids. Sequence analysis shows that PYBV is closely related to Potato virus A (PVA), with an overall 72% identity at the nucleotide level for the whole genome. The least conserved P1 protease gene shares only 50% nucleotide identity with PVA. The host range of PYBV was comparable to PVA on solanaceaous and non-solanaceous indicator plant species with the exception of Solanum demissum A and Y. Different symptoms were also observed for PYBV and PVA in Nicotiana benthamiana, Nicotiana hesperis and Nicotiana occidentalis P1. The susceptibility of potato (Solanum tuberosum) cultivars to PYBV and PVA was similar. In over 5 years of investigation, PYBV has not been found in commercial seed and ware potato crops in Scotland.     

Host range and some properties of Physalis mosaic virus,a new virus of the turnip yellow mosaic virus group

An apparently undescribed virus was isolated fromPhysalis subglabrata in Illinois, USA, and its properties were studied. The virus was namedPhysalis mosaic virus (PMV). It was readily transmitted by sap inoculation to 23 out of 34 Solanaceae tested, toChenopodium foetidum andSonchus oleraceus but not to 28 other non-solanaceous species inoculated. Purified preparations of PMV contained isometric particles of 27 nm in diameter, which sedimented as two components with sedimentation coefficients of 50 and 112 S. The 112 S component was infectious, the 52 S component was not. The virus contained 38% ribonucleic acid with a molar base content of G 14.4%, A 22.9%, C 37,2% and U 25.5%.Purified preparations were highly infectious; a concentration of about 6000 particles per ml was infectious on plants.PMV is a member of the Andean potato latent virus subgroup of the turnip yellow mosaic virus group. The virus was closely related to the viruses: Andean potato latent, belladonna mottle, dulcamara mottle and egg-plant mosaic.Samenvatting Een nog niet eerder beschreven virus, dat in de staat Illinois (V.S. van Amerika) opPhysalis subglabrata was gevonden, werd in Wageningen bestudeerd. Het virus dat Physalis mosaic virus (PMV) (in het Nederlands:Physalis-mozaïekvirus) werd genoemd, kon met sap worden overgebracht.BehalveChenopodium foetidum enSonchus oleraceus bleken ook 23 van de 34 getoetste soorten uit de familie Solanaceae vatbaar voor dit virus te zijn. Gezuiverde virus preparaten bevatten isometrische deeltjes met een diameter van 27 nm (Fig. 2) Het virus bestaat uit twee deeltjes met sedimentatie-coëfficiënten van 112 en 50 S. Het 112 S deeltje bleek infectieus te zijn, het andere niet. Op grond van de sedimentatiecoëfficiënten kan worden berekend dat het 112 S deeltje 38% nucleïnezuur bevat. Voor de basenverhouding in het nucleïnezuur werd 22,9% adenine, 14,4% guanine, 37,2% cytosine en 25,5% uracil gevonden (Tabel 1). Het hoge gehalte van cytosine kwam ook tot uiting in de U.V. absorptiekromme van het virus en het nucleïnezuur (Fig. 1). Het gezuiverde virus bleek zeer infectieus te zijn; 6000 deeltjes/ml waren in staat een plant van de soortNicotiana clevelandii ziek te maken.Op grond van serologisch onderzoek kon het virus tot de turnip yellow mosaic virus groep worden gerekend. Het vertoonde serologische verwantschap met de Andean potato latent virus (APLV) subgroep (Tabel 2). In premunitieproeven bood het slechts een geringe bescherming tegen APLV en dulcamara mottle virus. Het omgekeerde werd eveneens geconstateerd. De leden van de APLV-subgroep kunnen op grond van hun waardplantenreeks van elkaar onderscheiden worden (Tabel 3).     

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.     

广西部分烟区烟草曲叶病病原的分子鉴定

[目的]明确广西西部地区靖西(JX)、凌云(LY)、德保(DB)和乐业(LeY)等4个县市烟草曲叶病的病原。[方法]2010年5-6月分别从广西靖西、凌云、德保和乐业等县市采集具有典型曲叶症状的烟草叶片,用基于双生病毒DNA保守序列设计简并引物Bego-1和Bego-6对病叶组织总DNA抽提物进行PCR扩增和对PCR产物进行序列测定,用BLAST、Vector NTI、MEGA 4.0和Simplot program 3.2软件等进行病毒序列分析、系统进化树构建和病毒重组分析。[结果]从选取的9个表现典型曲叶症状的样品叶组织总DNA抽提物中均可扩增出约1500bp与预期大小相符的DNA片段。测序和序列比对分析显示,9个样品扩增产物核苷酸序列相似性为73.7%～99.2%,与已报道的双生病毒具较高的相似性。其中,JX-2与中国番茄曲叶病毒广西番茄分离物(G32)的相似性最高,达99.2%;JX-3和JX-5与云南胡椒曲叶病毒云南辣椒分离物(YN323)相似性最高,分别为92.5%和93.4%;LeY-1、LY-1、DB-1、JX-1、JX-4和JX-6则与中国番茄黄化曲叶病毒中国番茄分离物(CHI)和广西烟草分离物(G102)的相似性最高,均高于95.0%。基于PCR扩增产物及已报道的双生病毒属代表种相应核苷酸序列构建的系统进化树分析表明,9个广西烟草分离物分属3个簇群:中国番茄曲叶病毒簇、云南辣椒曲叶病毒簇和中国番茄黄化曲叶病毒簇。重组分析结果表明:JX-3是云南辣椒曲叶病毒和中国番茄曲叶病毒的重组病毒,JX-5是云南辣椒曲叶病毒和中国番茄黄化曲叶病毒的重组病毒。[结论]9个广西烟草分离物分属于4种双生病毒:中国番茄曲叶病毒和中国番茄黄化曲叶病毒,以及分别由上述两种病毒与云南辣椒曲叶病毒重组而来的2种重组病毒。其中,中国番茄曲叶病毒自然侵染烟草、云南辣椒曲叶病毒和中国番茄曲叶病毒及中国番茄黄化曲叶病毒的重组病毒等结果此前均未见报道。     

Evaluating aggressiveness and host range of Alternaria dauci in a controlled environment

The aggressiveness of Alternaria dauci isolates was investigated in greenhouse conditions. Twenty‐seven isolates were pre‐selected from a large collection to represent high diversity according to geographic or host origins and intergenic spacer (IGS) polymorphism. IGS sequence analysis revealed that isolates were grouped within three different clusters. Eleven isolates were selected and inoculated on a susceptible carrot cultivar. Three criteria (mean lesion number, mean necrotic leaf area and mean disease index) were used to assess the aggressiveness of isolates. Continuous variation in aggressiveness was shown and no clear division into isolate classes was evident. For the host range study, two isolates were inoculated under greenhouse conditions onto nine cultivated Apiaceae species, two wild Daucus species and six cultivated non‐Apiaceae species representing six botanical families. Lesions varying in severity were observed on all dicot species (Apiaceae and non‐Apiaceae), but no symptoms developed on the two monocots studied (leek and sweetcorn). Plant species were also differentiated on the basis of expanding lesions (cultivated and wild carrot, dill and fennel) or non‐expanding lesions (other dicot species). Typical A. dauci conidia were observed after in vitro incubation of leaves with symptoms. Fungal structures were isolated from lesions and A. dauci was confirmed on the basis of conidial morphology and specific conventional PCR results. Genotyping of individual isolates performed with microsatellite markers confirmed the presence of the inoculated isolate. The results clearly showed that, in controlled conditions, the host range of A. dauci is not restricted to carrot.     

<Emphasis Type="Italic">Mycosphaerella graminicola</Emphasis> produces a range of cell wall-degrading enzyme activities <Emphasis Type="Italic">in vitro</Emphasis> that vary with the carbon source

In this study, two Mycosphaerella graminicola isolates produced a range of cell wall-degrading enzymes (CWDE) in vitro that can potentially degrade wheat cell walls. The influence of three carbon sources on CWDE in vitro production was tested: 1) 1 % galactose (w/v), 2) 1% wheat cell walls (w/v) and 3) a mixture of 1% galactose (w/v) and 1% wheat cell walls (w/v). Six major activities produced by both isolates were detected: xylanase, β-1,3-glucanase, polygalacturonase, cellulase, β-xylosidase and β-galactosidase. Time-course experiments showed that different levels of enzyme activities were obtained with isolates 323 and 94269. These activities levels varied also with the type of carbon source used. Principal Component Analysis showed that the enzyme activities are gathered into two groups. None of the activities of the first group was correlated to the activities of the second group. It also showed that the optimal medium that allowed the production of most of the major activities contained both galactose and wheat cell walls.     

Lonicera latent virus,a new carlavirus serologically related to poplar mosaic virus: some properties and inactivation in vivo by heat treatment

A virus with elongate particles (656 nm) was isolated from severalLonicera species. This virus, apparently belonging to the carlavirus group, is serologically distantly related to shallot latent virus and closely related to poplar mosaic virus. The inability to infect poplar and two other hosts of poplar mosaic virus characterizes the virus fromLonicera as a new virus which was namedLonicera latent virus.The virus was easily sap-transmissible but was not transmitted byMyzus persicae.Dilution end-point was about 10^–3, thermal inactivation between 65°C and 80°C and ageing in vitro 1–6 days.Heat treatment, combined with tip-rooting appeared to be a good method to eliminate the virus from severalLonicera species and cultivars.Samenvatting In verschillende soorten en cultivars van het geslachtLonicera (kamperfoelie) blijkt een virus voor te komen dat gemakkelijk door sapinoculatie kan worden overgebracht op kruidachtige planten.Een tegen gezuiverd virus bereid antiserum had een titer van ca. 4096. Er kon mee worden aangetoond dat het virus van kamperfoelie serologisch nauw verwant is met populieremozaïekvirus (Tabel 1). Het virus van kamperfoelie is echter niet in staat om populier,Phaseolus vulgaris Bataaf enVigna sinensis te infecteren en wordt mede daarom als een afzonderlijk virus beschouwd. Het wordt aangeduid als latent kamperfoelievirus (Lonicera latent virus) en behoort evenals populieremozaïekvirus tot de carlavirusgroep (aardappelvirus-S-groep).Het virus blijkt vrij gemakkelijk te kunnen worden geëlimineerd door besmette kamperfoelieplanten gedurende ongeveer zes weken een warmtebehandeling (37°C) te geven en daarna de uiterste toppen (1 cm) te stekken. Van verschillende cultivars werd op deze wijze virusvrij uitgangsmateriaal verkregen.     

南宁市朱槿曲叶病毒病病原分子鉴定和寄主范围研究

［目的］ 明确南宁市朱槿曲叶病毒病的病原及其寄主范围,初步了解各朱槿品种的抗病性。［方法］ 对采自南宁市区的朱槿病株分离物进行分子鉴定及寄主范围测定;进行朱槿曲叶病毒病发生情况调查,根据发病率高低和症状轻重对南宁市常见的朱槿品种抗病性进行初步评价。［结果］ 基因克隆与序列比较结果表明,该病的病原为木尔坦棉花曲叶病毒（Cotton leaf curl Multan virus, CLCuMV）,其DNA A全长含2 737个核苷酸,伴随有卫星DNAβ分子,分子大小为1 346个核苷酸;研究发现该病毒可以通过B型烟粉虱传播,烟粉虱接种和PCR检测试验〖JP2〗结果表明,该病毒可侵染朱槿、红麻、棉花、西菲葵、黄蜀葵等5种供试的锦葵科植物,表现典型曲叶症状;初步调查评价结果,‘朱砂红朱槿’、‘泰国黄朱槿’、‘重粉朱槿’、‘锦球朱槿’和‘马旦朱槿’属于高度感病品种;‘阿美丽坚朱槿’、‘乳斑朱槿’属于感病品种;‘大红朱槿’、‘大红花’和‘粉喇叭’属于中抗品种;‘红龙朱槿’、‘红喇叭’、‘青杆吊钟’和‘七彩朱槿’属于高抗品种。［结论］ 上述结果可为有效控制朱槿曲叶病毒病提供依据。     

Molecular phylogenetic,morphological, and pathogenic analyses reveal a single clonal population of Septoria lycopersici with a narrower host range in Brazil

Septoria leaf spot, caused by Septoria lycopersici, is considered one of the most important diseases of tomato in Brazil. Despite its importance, the disease agent is still poorly studied. Septoria isolates collected from different production regions of Brazil were characterized by molecular, morphological, and pathogenic methods. A set of 104 isolates was sequenced for the DNA Tub, Cal, and EF1-α loci. Ten isolates were selected, according to geographical region of origin and type of leaf lesion (typical or atypical), for morphological characterization and for evaluation of aggressiveness on tomato cultivar Santa Clara. To evaluate the pathogen host range, cultivated and wild Solanaceae plants were inoculated with four selected isolates. The results showed that all isolates grouped with the type isolate of S. lycopersici in maximum likelihood and Bayesian inference trees. The isolates were morphologically similar. All isolates selected for pathogenicity testing on tomato were able to induce typical symptoms of the disease, but differed in their aggressiveness. A total of eight species of Solanaceae were also identified as potential alternative hosts for S. lycopersici. This information will provide a more accurate assessment of the risks involved with the introduction of new crops, especially of the genus Solanum, in areas where the species is already present. In addition, it will provide the basis for the establishment of more efficient methods in the management of Septoria leaf spot of tomatoes in natural conditions and in the different production systems.     

番茄黄化曲叶病毒V 2基因原核表达及多克隆抗体制备

 通过PCR的方法,从番茄黄化曲叶病毒江苏分离物DNA中扩增到V2基因,并克隆到pMD18-T载体,序列测定结果显示其与报道的XH2分离物序列完全一致,核苷酸序列全长351 bp,编码115个氨基酸,推测分子量约13 kD。将该V2基因亚克隆到原核表达载体PET32a中获得重组表达载体PET32a-V2,转化大肠杆菌BL21（DE3）,IPTG可诱导1个分子量约为32 kDa的融合蛋白（含His标签）表达,经Ni⁺ NTA亲和柱纯化,获得纯化的重组蛋白,免疫家兔制备TYLCV-V2蛋白的多克隆抗体Anti-V2,间接ELISA测定Anti-V2的效价达1∶655 360,Western blot及DIBA分析结果表明Anti-V2可与V2蛋白发生特异血清反应,可为进一步研究V2蛋白的功能提供参考。     

Characterization and pathogenicity of Septoria gaurina associated with leaf blotch disease on Gaura parviflora in China

Gaura parviflora, a flowering plant of the family Onagraceae, is regarded as an invasive plant in China, but is subject to leaf blotch disease. Based on morphological features, we identified the causal agent to be Septoria gaurina. However, the phylogenetic position of S. gaurina remains unknown. Multilocus phylogenetic analysis using the internal transcribed spacer (ITS), β-tubulin (Tub2), actin (Act), calmodulin (Cal), translation elongation factor 1-α (EF), and 28S nrDNA (LSU) nucleotide sequences indicated that the isolates belonged to the Septoria genus. Pathogenicity tests demonstrated that the isolates produced symptoms on the leaves of G. parviflora. These symptoms included small, water-soaked, circular lesions formed on leaves inoculated with conidial droplets, and bigger, water-soaked lesions on leaves inoculated by conidial spraying. To our knowledge, this is the first report of S. gaurina as the causative agent of leaf blotch disease of G. parviflora in China.     

Tomato leaf curl geminivirus in Australia: occurrence, detection, sequence diversity and host range

The occurrence of whitefly transmitted geminiviruses in Australia was studied using a mixed DNA probe capable of detecting a range of distinct geminiviruses. The only geminivirus species detected was Tomato leaf curl virus (TLCV), which is spread across a vast geographical region of far-northern coastal Australia, an area inhabited by the Australasian-Oceania biotype of Bemisia tabaci . The newly introduced silverleaf whitefly, B. tabaci biotype B, forms high population densities in the eastern coastal region of Queensland and is currently located approximately 150 km from the nearest known TLCV-infected area. The viral host range appeared to be narrow and of 58 species of crop plants and weeds inoculated using the B biotype, only 11 became infected with the virus, including five that did not show foliar symptoms. A DNA fragment of 694 nt, including the complete C4 open reading frame (ORF), the overlapping N-terminal part of the C1 ORF and the viral iterons involved in replication, was amplified from 11 TLCV field isolates and sequenced. Sequence analysis revealed an overall sequence variation of up to 14% in this region, as well as the presence of distinct viral iterons.     

Rice yellow mottle,a mechanically transmissible virus disease of rice in Kenya

In Kenya around Lake Victoria rice is affected by a hitherto undescribed virus for which the name Rice Yellow Mottle Virus (RYMV) is proposed. The virus was easily transmitted mechanically to rice and toOryza barthii andOryza punctata, but not toOryza eichingeri, barley, bulrush millet, durum wheat, finger millet, maize, oats, rye, sorghum, wheat, sugarcane, 20 other monocotyledonous and 9 dicotyledonous plant species.The disease is characterised by stunting and reduced tillering of the rice plant, crinkling, mottling and yellowish streaking of the leaves, malformation and partial emergence of the panicles, and sterility. In severe cases the plant may die. RYMV is stable and highly infective.The vector is the beetleSesselia pusilla Gerstaeker (fam. Chrysomelidae, Galerucinae). The beetle has been identified by Mr John A. Wilcox, New York State Museum and Science Service, New York.S. pusilla transmitted the virus for at least five successive days after feeding on an infective plant.Purified virus preparations consisted of polyhedral particles about 32 m in diameter. The sedimentation coefficient was 116S.Samenvatting In Kenya wordt rijst, voornamelijk Sindano, verbouwd op familiebedrijfjes aan de kust van de Indische Oceaan en langs het Victoria Meer, en door pachters van het meer naar het binnenland gelegen Mwea Irrigation Settlement. In de omgeving van Kisumu (Nyanza Province), waar rijst geteeld wordt volgens slechte cultuurmethoden, wordt sinds 1966 een tot dusver onbekende ziekte, hier rice yellow mottle genoemd, waargenomen.In het veld vallen de zieke planten onmiddellijk op door de gelige verkleuring van de bladeren. De jongste bladeren zijn gevlekt of geel-groen gestreept (Fig. 1). De uitstoeling is gering en de plant gedrongen (Fig. 2). De zaadopbrengst wordt sterk verlaagd door het optreden van steriliteit.Ongeveer 7 dagen na sap-inoculatie van Sindano-zaailingen verschijnen de eerste symptomen. Na inoculatie in een jong stadium van de plant krullen de eerstgevormde bladeren spiraalvormig (Fig. 3). Later worden de bladeren geel en necrotisch. Jong geïnfecteerde planten kunnen zelfs afsterven. De pluimen komen onvoldoende uit de bladschede, zijn misvormd en dragen veelal kleine en misvormde bloembakjes, die meestal loos zijn (Fig. 4). Sindano rijst, 3 weken voor het in pluim schieten geïnoculeerd, liet een zeer sterke opbrengstvermindering zien.In sap, verkregen van wortels van zieke rijstplanten, in de guttatievloeistof en in irrigatiewater van ernstig zieke rijstvelden kon het virus gemakkelijk worden aangetoond. Overdracht van RYMV door zaad en grond werd niet waargenomen.RYMV was nog infectieus na een verhitting gedurende 10 min bij 80°C. De verdunningsgrens hing af van het tijdstip waarop het bladmateriaal van de zieke planten getoetst werd en liep uiteen van 10^–10 (2–3 weken na inoculatie) tot 10^–6 (4–5 weken na inoculatie). Sap bewaard bij kamertemperatuur was nog infectieus na 33 dagen, maar had deze eigenschap verloren na 51 dagen. Bewaard in een koelkast bleef het sap zeker 71 dagen infectieus.A1 de getoetste rijstrassen werden na sap-inoculatie door het virus geïnfecteerd, evenalsOryza barthii enOryza punctata (Fig. 5). Niet vatbaar waren:Oryza eichingeri, baardtarwe, eleusine gierst, gerst (Proctor), haver (M.F.C. 15/67 en Lampton), mais (Hybrid 611B, Hybrid 612 en Hybrid 613B), parelgierst, rogge, sorghum (H726 en H6060), suikerriet (NCo 310 en Q 45), tarwe (Kenya Kudu en Wisconsin), evenals 20 andere monocotyle en 9 dicotyle plantesoorten, waaronder de doorgaans bij plantevirusonderzoek gebruikte toetsplanten.Het kevertjeSesselia pusilla Gerstaecker (fam. Chrysomelidae, Galerucinae; zie Fig. 6) bracht het virus over. Een enkel insekt was in staat het virus gedurende tenminste 5 dagen over te brengen.RYMV was gemakkelijk te zuiveren. Hierbij werd uitgegaan van 16,5–20 g jonge duidelijk zieke bladschijven van jonge 2–3 weken tevoren geïnoculeerde rijstplanten (Sindano) door uitschudden met chloroform en uitzouten met ammoniumsulfaat, iets gewijzigd naar Proll en Schmidt (1964). De elektronenmicroscopische beelden van de gezuiverde virussuspensie laten één type veelkantige deeltjes zien met een diameter van ongeveer 32 m (Fig. 7). De S₂₀-waarde is 116S (Fig. 8 en 9).Ter bestrijding van de ziekte moet op korte termijn allereerst de cultuurmethode van de rijst op de familiebedrijfjes verbeterd worden. Het gehele jaar in verschillende groeistadia voorkomen van rijst op een klein gebied en veelal zieke opslag op juist geoogste velden, dragen belangrijk bij tot de verspreiding van de ziekte. Het gebruik van kortgroeiende rijst-varieteiten wordt aanbevolen om overlapping van de groeiseizoenen te voorkomen.This paper is published with permission of the Chief of Technical Services, Ministry of Agriculture, Kenya.     

甘蔗黄叶病及其病原分子生物学研究进展

甘蔗黄叶病是由甘蔗黄叶病毒(Sugarcane yellow leaf virus,SCYLV)引起的一种病毒病害,在全球主要甘蔗种植国家或地区普遍发生,危害日益严重。SCYLV经甘蔗蚜虫以持久性方式传播,在寄主植株内主要侵染韧皮部组织。该病毒起源于黄症病毒科属间基因重组,被国际病毒分类命名委员会确认为黄症病毒科马铃薯卷叶病毒属成员。文章综述了甘蔗黄叶病毒生物学特征、病害发生和危害、病原鉴定和检测、分子进化和遗传多样性、基因组结构和基因功能以及抗病转基因等方面研究进展,并对甘蔗黄叶病抗病育种和防治措施作了讨论。     

河北省番茄黄化曲叶病毒和番茄褪绿病毒复合侵染及分布

 番茄黄化曲叶病毒(Tomato yellow leaf curl virus, TYLCV) 及番茄褪绿病毒(Tomato chlorosis virus, ToCV)是危害番茄的主要病毒。2016~2017年,在河北省18个番茄主产区调查番茄病毒病危害情况,采集262份植株矮化、叶片黄化、褪绿、卷曲的番茄样品,利用分子生物学技术对病原进行鉴定。结果表明:2016~2017年河北省番茄病毒病发生普遍,所检样品中TYLCV的侵染率为68.66%,其中与ToCV复合侵染率为19.5%;除栾城、滦南、乐亭、永年4地样品暂未检测到ToCV外,其他14个采样点的样品均检测到ToCV,侵染率为19.5%,且全部表现为与TYLCV复合侵染,河北省番茄主产区暂未发现ToCV单独侵染的样品。     

Pathogenicity,host range and activities of a secondary metabolite and enzyme from Myrothecium roridum on water hyacinth from Thailand

Leaf blight disease of water hyacinth was observed and collected from different geographical areas of Thailand. The disease is caused by a fungal pathogen that was identified as Myrothecium roridum by using its morphological characteristics. The most effective fungal strains were evaluated for pathogenicity on water hyacinth under greenhouse and natural conditions. Myrothecium roridum isolate, Kamphaeng Sean Campus (KKFC) 448, was found to be the most virulent. Different fungal formulations were evaluated for their level of control of water hyacinth. The results showed that spore suspensions with 10% palm oil or 1% Tween 20 caused a higher level of disease severity, compared to spores applied in water alone. The host range of KKFC 448 was evaluated by using 77 plant species that belong to 40 plant families. The fungus did not cause disease on 74 economically important plants but did produce disease signs on water hyacinth and two other aquatic weeds, duckweed and water lettuce. Leaf blight occurs on water hyacinth leaves after being treated with crude extracts of M. roridum and it was indicated that secondary metabolites were released from the fungal mycelia. Myrothecium roridum that was grown on boiled paddy rice produced β‐1,4‐exoglucanase, β‐1,4‐endoglucanase, β‐glucosidase, xylanase and pectinase more than Mroridum that was grown on potato dextrose agar. The results indicated that M. roridum is a pathogen of water hyacinth and the fungus is capable of producing different enzymatic activities on potato dextrose agar and boiled paddy rice, which might be important for infection.