Abiotic transmission of Rice yellow mottle virus through soil and contact between plants

The roles of guttation fluid, irrigation water, contact between plants and transplantation into contaminated soil in the transmission of Rice yellow mottle virus (RYMV) were assessed. RYMV presence and infectivity were tested by Enzyme-Linked Immunosorbent Assay (ELISA) and by inoculation to susceptible rice cultivar BG90-2. The virus was readily detected in guttation fluid collected from infected rice plants. Transmission tests from this fluid led to high disease incidence (86.6%). Irrigation water collected at the base of infected plants growing in pots was less infectious, as inoculations led to disease incidences below 40%. No virus was detected and could be transmitted from field-irrigation water. Up to 44% healthy rice plants whose leaves were in contact with those of infected plants became infected but, no transmission occurred through intertwined roots. Transplantation of rice seedling into virus-contaminated soil also led to plant infection. However, virus survival in the soil decrease rapidly and infectivity was completely lost 14 days after soil contamination. Altogether, these results indicated that high planting densities of rice are likely to favour secondary spread of rice yellow mottle disease. Transplantation of rice seedlings not earlier than 2 weeks after soil preparation should prevent soil transmission of the virus. Although guttation fluid is highly infectious its contribution to virus infectivity in irrigation water is negligible as field-irrigation water was not found to be an infectious source for RYMV.     

Transmission of Rice Black-Streaked Dwarf Virus from Frozen Infected Leaves to Healthy Rice Plants by Small Brown Planthopper (Laodelphax striatellus)

In order to preserve virus for identifying the resistance of rice varieties against rice black-streaked dwarf disease, a simple and reliable method was developed, through which virus-free small brown planthopper (SBPH) acquired rice black-streaked dwarf virus (RBSDV) from frozen infected leaves and the virus was transmitted to healthy rice plants. The experimental results showed that SBPH could obtain RBSDV from frozen infected rice leaves and the virus could be transmitted to a susceptible rice variety. For the ability to acquire RBSDV and transmit the virus to healthy plants by SBPH, there was no significant difference between frozen infected leaves and in vitro infected leaves. The novel method could be applied to identification of rice variety resistance to rice black-streaked dwarf disease, facilitating the breeding process for rice black-streaked dwarf disease resistance.     

THE SIGNIFICANCE OF VIRUS DISEASES FOR THE PRODUCTIVITY OF GRASSLAND

The effect of cocksfoot streak virus (CSV) on Dactylis glomerata L. and barley yellowdwarf virus (BYDV) on Lolium perenne L. was investigated under both single-plant and simulated-sward conditions. The effect of cocksfoot mottle (CFMV) was examined on single plants. CFMV was lethal, and in conditions of inter-plant competition CSV was semi-lethal and BYDV was non-lethal. When the virus was lethal, a considerable lag-phase was observed between mortality and complete compensation in yield. When the virus was non-lethal, plant growth was restricted, either horizontally (CSV) or vertically (BYDV). In mixed, healthy and infected plant populations healthy individuals compensated only for reductions in lateral growth.
CSV infection transformed plants of a pasture variety into 'hay' types and BYDV infection transformed plants of a hay variety into 'pasture' types. CSV increased the plant's aggressiveness in hay conditions, but reduced its ability to withstand frequent defoliation. Conversely, BYDV increased the plant's aggressiveness in pasture conditions, but reduced it in hay conditions. Compensation from healthy plants occurred only in CSV-infected swards subjected to frequent defoliation. Some possible reasons for the absence of compensation in other treatments are discussed.     

建兰花叶病毒CP基因的原核表达及抗血清制备

通过间接酶联免疫检测和电镜观察对从福建省漳州市采集的卡特兰病样进行检测,证明样品感染了建兰花叶病毒。设计一对特异性引物,扩增并克隆病毒分离物的外壳蛋白基因,随后将目的基因插入pET-29a（＋）中构建相应的原核表达载体。目的蛋白经诱导表达及纯化后免疫家兔并获得了特异性抗血清。Westernblot检测结果表明,抗血清与诱导表达的CyMV外壳蛋白发生特异性反应。间接酶联免疫法检测结果表明,抗血清可检测病汁液的最低稀释度达1∶51200,最佳工作浓度为1∶1000,病汁液灵敏度为0.39mg/mL,而与TMV等11种同源或异源病毒均无明显的血清学交叉反应。     

Detectability and translocation of potato virus A in potato plants after artificial inoculation

Summary The tip leaflets of median leaves of plants of different ages of the very susceptible cv. Jara and the less susceptible cv. Krasava were dry inoculated with potato virus A (PVA). Movement of the virus was followed by using the A6 leaf test on different plant perts at intervals after inoculation. The technique was not very reliable but the results showed that in cv. Jara the rate of movement of virus within the plants and into the tubers was greater and less dependent on their age than in the less susceptible cv. Krasava. In cv. Jara. PVA reached the tubers with 6–7 days from inoculation but in cv. Krasava not until after 21 days and not at all if the plants were more than 6 weeks old when inoculated.     

Comparative defoliation tolerance of temperate perennial grasses

The defoliation tolerance of cultivars of four temperate perennial pasture grasses, perennial ryegrass (Lolium perenne, cv. Yatsyn1), phalaris (Phalaris aquatica cv. Australian), tall fescue (Festuca arundinaceae cv. Demeter) and cocksfoot (Dactylis glomerata cv. Porto), was determined under controlled conditions over a period of 12 weeks. Undefoliated plants were compared with defoliated plants, where only half of one leaf was left intact at the initial defoliation, and leaf regrowth was harvested every 3–4 d. The growth responses measured were plant tiller number, dry weight, relative leaf regrowth rate, root:shoot ratio, sheath:stem ratio and specific leaf weight. All species showed morphological adaptations that potentially increased their ability to tolerate defoliation (e.g. increased allocation to shoot at the expense of roots and lower specific leaf weights) but cocksfoot was found to be the most defoliation‐tolerant and perennial ryegrass the least. The adaptation that favoured cocksfoot most strongly was high sheath:stem ratio, which, it is proposed, allowed it to maintain photosynthesis and a level of carbon supply sufficient to support regrowth throughout the experiment. The strategy of perennial ryegrass which favours leaf growth and leads to rapid leaf turnover rates made it particularly susceptible to defoliation under the conditions of this experiment. This highlights the likely importance of defoliation‐avoidance responses in explaining the well‐known grazing resistance of this species. Phalaris and tall fescue showed responses that were intermediate between the other two species. The importance of defoliation‐avoidance mechanisms and implications for grazing management are discussed.     

Host genes and transgenes that confer resistance to a Scottish isolate of potato leafroll virus are also effective against a Peruvian isolate

Summary Potato genotypes with host gene-mediated resistance (host-MR) and coat protein-mediated transgenic resistance (CP-MR) to potato leafroll virus (PLRV), were inoculated with a Scottish and a Peruvian isolate of PLRV. The coat protein transgene had been cloned from the Scottish PLRV isolate which had also been used during the screening and selection of genotypes with host resistance. Significantly less PLRV accumulated in plants with either host-MR or CP-MR than in plants of susceptible genotypes or in non-transformed control plants, but the two forms of resistance were equally effective against both PLRV isolates.     

Transmission of potato leafroll virus to or from tuber slices by aphids feeding through a membrane

Summary Eye-bearing slices, cut from healthy potato tubers and placed between Parafilm membranes, were inoculated with potato leafroll virus (PLRV) byMyzus persicae. PLRV was detected by ELISA and by transmission tests in tuber slices and in plants grown from the slices of the susceptible cv. Désirée, but not in those of the resistant cv. Arkula. These results suggest that PLRV replication and transport within tuber phloem is controlled by specific mechanisms of resistance.M. persicae was also able to acquire and transmit PLRV toPl floridana from slices cut from tubers of infected plants. The aphids effectively transmitted PLRV from slices cut from the sprouting rose end but they failed to transmit it from slices cut from the heel end of tubers.     

EAR-BEARING CAPACITY OF TILLERS ARISING AT DIFFERENT TIMES IN HERBAGE GRASSES GROWN FOR SEED

Three experiments are described, two with S215 meadow fescue and one with S37 cocksfoot, all grown for seed, during which all new tillers, soon after their appearance, were labelled at intervals of a month—a total of nearly 12,000 tillers. Whatever the first date of observation, the earliest tillers were always found to have contributed the largest proportion of the ears present at harvest. In one experiment applied nitrogen caused an increase in the number of tillers and ears, without however affecting the percentage contribution of the earliest tillers to the final crop of ears. The importance of the autumn period of tillering is stressed.     

ENVIRONMENTAL EFFECTS ON SYMPTOM EXPRESSIONS OF VIRUS DISEASES IN GRASSES

Viruss damage was observed on plants of Lolium perenne L. and Dactylis glomcrata L. undergoing growth-analysis studies. On the basis of visual symptoms the damage was presumed to be caused by barley-yellow-dwarf virus (BYDV) in ryegrass and by streak virus (CSV) in cocksfoot. The number of plants which could be clinically diagnosed as infected and the degree of damage were both positively correlated with temperature. It was inferred that at low temperatures both viruses could remain relatively inactive, or even latent. This was subsequently confirmed by removing cocksfoot plants from a heated glasshouse (19°–23°C) to a cold environment (6–10°C) which resulted in a progressive loss of symptoms in newly developing leaves. Some possible explanations and practical implications are briefly discussed.     

SEASONAL TILLER POPULATIONS OF GRASS AND GRASSJCLOVER SWARDS WITH AND WITHOUT IRRIGATION

The number of tillers per unit area of pure swards of S23 and S24 perennial ryegrass, S37 cocksfoot, S48 timothy and of a S23 perennial ryegrass/white clover sward, were examined at intervals of 4 weeks during a period of two years. The monthly appearance of new tillers and the death of tillers of S23 and S24 ryegrass and S48 timothy were followed for 15 months. The studies were made with and without irrigation.
The number of tillers varied with season through the year. Tiller numbers increased rapidly in late winter or early spring, and then declined until midsummer. Numbers rose again between June and August. This seasonal pattern of tiller numbers was most marked in ryegrass and timothy and least in cocksfoot. The effect of irrigation on tiller numbers was variable, and usually small in relation to the overall seasonal fluctuations in tiller numbers, but dry soil conditions could delay an increase in tiller numbers between June and August. The relationship between seasonal fluctuations in tiller number and root growth of swards is discussed.     

灰飞虱从冷冻病叶获得水稻黑条矮缩病毒方法的研究初报

  研究出一种利用灰飞虱从冷冻病叶中获得水稻黑条矮缩病毒（rice black streaked dwarf virus, RBSDV）的方法。以-70℃条件下冷冻保存的RBSDV罹病植株叶片对灰飞虱饲毒,随后接种感病水稻品种,结果发现灰飞虱可从冷冻病叶上获得RBSDV,并传播RBSDV,且获毒能力和传毒能力与新鲜病叶没有差异。这表明从冷冻病叶上获得RBSDV的灰飞虱可以应用于品种抗性鉴定,此法可望加快RBSDV抗病品种的选育进程。     

番茄斑驳花叶病毒海南分离物全基因组序列分析

本研究采用小RNA深度测序及RT-PCR检测技术鉴定海南省陵水县冬种番茄(圣女果)病株受到烟草花叶病毒属的番茄斑驳花叶病毒(Tomato mottle mosaic virus,ToMMV)侵染。通过机械摩擦接种,将该病原病毒保存于本生烟植株上,命名为番茄斑驳花叶病毒海南分离物(ToMMV-HN)。采用RT-PCR分段扩增获得了该病毒分离物的全基因组序列。测序结果表明,该病毒分离物基因组全长6399 nt,含有4个开放阅读框,编码4种蛋白,与已报道的ToMMV9个分离物核苷酸和编码产物氨基酸序列相似性分别为98.0%～100%和99.2%～100%;与同属的番茄花叶病毒(Tomato mosaic virus,ToMV)、番茄棕色皱果病毒(Tomato brown rugose fruit virus,TBRFV)和烟草花叶病毒(Tobacco mosaic virus,TMV)代表分离物核苷酸序列相似性分别为84.7%、80.7%和79.0%。系统进化分析结果表明,ToMMV-HN与该病毒中国西藏和云南辣椒分离物亲缘关系最近,与美国加州和南卡州番茄分离物次之,与西班牙和巴西番茄分离物亲缘关系相对较远。本研究为我国番茄病毒病诊断和防控,尤其是抗病品种选育提供了基础资料。     

Sweetpotato viruses in China

China is the largest sweetpotato producer country in the world, with its total growing area and yield reaching 5.5 million ha and 106 million metric tones, respectively. Viral diseases constitute a major hindrance to the development and highly profitable production of the sweetpotato industry. The present article provides updated comprehensive information on type of virus, yield loss caused by viruses, increased yield and benefits by cultivation of virus-free plants. By analyzing data on changes in vegetative growth and physiology of plants infected by viruses, the authors attempted to elucidate the mechanisms by which infection of viruses caused significant losses of the root tuber yield. Meristem culture techniques for production of virus-free sweetpotato plants and propagation system of virus-free seed tubers have been well developed in China. At present time, virus-free sweetpotato seed tubers have been widely used in major sweetpotato-growing areas in China. Usage of virus-free seed tubers has brought great benefits to the sweetpotato industry in China.     

Beitrag zur Bestimmung der relativen Resistenz gegen Potato Virus X (PVX) an In-vitro-Pflanzen der Kartoffel

Zusammenfassung In-vitro-Pflanzen von PVX-resistenten (‘Schwalbe’, ‘Karella N’) und--anf?lligen Genotypen (‘Turbella N’, ‘Ogonjok’) in unsteriler Kultur auf Kiessubstrat wurden mechanisch inokuliert (Isolat PVX-‘Erstling’, Glasst?bchen). Eine quantitative Absch?tzung der relativen Viruskonzentration mittels ELISA 7...42 Tage nach der Inokulation (dpi) ergab bei den resistenten Genotypen im Mittel von 5 Versuchen mit 82 und 83% (70...90%) einen gegenüber den anf?lligen Formen (19 und 20%, 0...40%) signifikant h?heren Anteil Pflanzen mit gehemmter Virusvermehrung (E_{405 nm}≤1,0). Eine damit übereinstimmende Einstufung erfuhren die 4 geprüften Genotypen bei Absch?tzung der relativen Viruskonzentration an prim?rinfizierten Topf- und sekund?rinfizierten Freilandpflanzen.

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Summary The suitability of in vitro plants for determining relative (quantitative) PVX resistance was examined using resistant (‘Schwalbe’, ‘Karella N’) and susceptible (‘Turbella N’, ‘Ogonjok’) genotypes. They were grown in non-sterile gravel augmented with 2 ml of 0.45% Wopil solution (a mixture of nutrient salts with trace elements; VEB Chemiekombinat, Bitterfeld, DDR) and mechanically inoculated (glass rod) with the ‘Erstling’ isolate of PVX. The relative virus concentration in 10–15 in vitro plantlets of each genotype was estimated quantitatively using ELISA after 7, 14, 21, 28, 35 and 42 days (Fig. 1). The resistant genotypes had significantly higher proportions of plants with restricted virus multiplication (E_{405 nm}≤1.0) than the susceptible ones: means over five experiments were 82 and 83% for the resistant genotypes (70...90%) and 19 and 20% for the susceptible (0...40%)-Fig. 2. Estimation of the virus concentration in primary infected pot and secondary infected field plants of the four tested genotypes showed good agreement with established resistance rankings.

     

New disease symptoms observed on field-grown potato plants with potato spindle tuber viroid and potato virus Y infections

Summary Potato spindle tuber viroid (PSTV) and potato virus Y (PVY) were isolated from plants of cultivar Kennebec with severe necrotic symptoms in the field. In the greenhouse, severe necrotic symptoms were reproduced only when potato plants were infected either simultaneously with PSTV+PVY, or with PSTV prior to PVY infection. Thirteen additional potato cultivars were tested in the greenhouse for this synergistic reaction and eight developed necrotic responses similar to cv. Kennebec. PVY concentration was significantly higher in doubly infected plants, compared with those infected with PVY alone.     

The Rate of Virus Spread in New Potato Cultivars in the North of Poland

The aim of this research was to assess the rate of increase in the level of tuber infection by PVY, PVM, and PLRV during three consecutive years of multiplication in the field for 17 new cultivars registered in Poland in 2009–2011 and for two cultivars not listed in the registry but popularly cultivated. The research was conducted in Bonin near Koszalin (north-western Poland) in 2010–2013. Tuber infection was assessed using DAS ELISA in a grow-out test in the winter-spring period. Among the 19 cultivars examined, eight had high resistance to PVY (above grade 8 on a scale of 1–9); during the 3-year research period, they were not infected. Also, cv. Gawin seemed more resistant than previously assumed. In contrast, cvs. Hermes and Sylvana, which were rated in the Netherlands as quite resistant, were clearly very susceptible (grade 3–4) in Polish conditions, as within 2 years 100% of tubers were infected with this virus. The greatest susceptibility to PVM was shown by cvs. Danuta and Stasia, 50% of which were infected, despite moderate exposure to the virus. Cultivars Zenia, Etiuda, Jubilat, and Viviana appeared very resistant to PVM as the number of infected tubers did not exceed 5%. PVM was not detected in the tubers of cvs. Bursztyn, Gustaw, and Legenda, which confirmed that these cultivars possess the Rm gene. Not all cultivars regarded susceptible (grade 3–4) to PLRV were infected. The virus was not found in tubers of cv. Bursztyn, while in cv. Hermes, assessed in the Netherlands as being quite resistant, almost 40% of tubers were infected.     

The susceptibility of agricultural grasses to pre-emergence damage caused by Fusarium culmorum and its control by fungicidal seed treatment

A total of 104 cultivars of six agricultural grass species were compared under controlled conditions for their relative susceptibility to the common soil-borne fungus Fusarium culmorum (W.G.Sm.) Sacc. A number of the perennial ryegrass ( Lolium perenne ) cultivars showed a high degree of resistance, their emergence being little reduced under the stringent test conditions. The emergence of others was, however, severely affected. Italian ryegrass ( L. multiflorum ) cultivars generally had a high level of resistance, the outstanding exception being cv. Leda Daehnfeldt which proved very sensitive. None of the cultivars of cocksfoot ( Dactylis glomerata ), timothy ( Phleum pratense ), meadow fescue ( Festuca pratensis ) or tall fescue ( F. arundinaceae ) which were tested had more than a moderate level of resistance and most were highly susceptible.
The vigour of ryegrass seedlings was, in most cases, closely correlated with percentage emergence but was less closely correlated in the more susceptible species. These often had a very low emergence but those seedlings which did establish were frequently of a size similar to that of healthy plants.
Treatment of grass seed with a fungicide mixture of benomyl (15% a.i.) plus captan (60% a.i.) provided excellent protection against F. culmorum on even the most susceptible cultivars.
Drazoxolon (40% a.i.) was, however, much less effective.     

Tiller production in cocksfoot (Dactylis glomerata) and tall fescue (Festuca arundinacea) growing along a light gradient

Pasture managers seek to balance leaf appearance with utilization to sustain sward productivity while meeting livestock nutritional needs. Achieving this in silvopasture must account for the influence of light and defoliation on tillering. We determined tiller production, as a function of light availability and clipping height, for two forage grasses adapted to cool, humid temperate conditions of the Appalachian region of the USA. Tiller production of cocksfoot (orchard grass) (Dactylis glomerata L.) and tall fescue (Festuca arundinacea Schreb.) was greatest in unshaded and least in shaded sites, irrespective of residual clipping height. Juvenile plants produced more than twice as many tillers as mature plants, with differences accentuated by site, reflecting a shift in reproductive and resource‐use strategies. Trends in tiller production were as follows: cocksfoot > tall fescue; 10‐cm > 5‐cm residual clipping height; and unshaded > partially shaded > densely shaded sites. Tiller production reaches maximum earlier in the woodlot site, regardless of species or clipping height, than at the unshaded site. Plants at the wooded site extended leaves quickly, sustaining herbage production, but had less non‐structural carbohydrate and fewer tillers. Cocksfoot, but not tall fescue, sustained leaf and tiller production in the shaded sites, suggesting that cocksfoot is suited for use in silvopasture in this region.     

Production of phytoalexins, glycoalkaloids and phenolics in leaves and tubers of potato cultivars with different degrees of field resistance after infection withPhytophthora infestans

Summary The kinetics of accumulation of phytoalexins, glycoalkaloids and phenolics was studied in two potato cultivars differing in their degrees of field resistance when infected withPhytophthora infestans. Tuber slices and leaves of cvs Pampeana INTA (high degree of field resistance, free of R genes) and Bintje (susceptible) were infected with race C (complex race 1, 3, 5, 7, 11) ofPhytophthora infestans. Phytoalexins and phenolics accumulated in tuber and leaf tissues which had been inoculated. The levels of these compounds in the susceptible cv. Bintje were relatively low and similar to those found before inoculation. Leaves of cv. Pampeana INTA had a very high glycoalkaloid content, suggesting that glycoalkaloids may play a role in protection of leaves against the fungus. However, we could find no correlation between resistance and glycoalkaloid content of tubers. Our results suggest a major role of phytoalexins, phenolics and glycoalkaloids in the complex mechanisms of field resistance.