Survey for viruses infecting onion, garlic and leek crops in Iran

Surveys to identify virus diseases affecting garlic ( Allium sativum ), onion ( Allium cepa ) and Persian leek ( Allium ampeloprasum var. persicum ) were conducted from 1999 to 2002. Surveys covered different regions of Iran (Tehran [different vegetable markets, farmer fields and cultivation areas], Noushahr, Chalous, Roudbar, Sari, Hamadan, Touyserkan, Ghazvin and Jiroft). A total of 2045 (1285 garlic, 525 onion and 230 leek) samples showing symptoms of virus infection were collected and tested by ELISA; and in some cases tests were also confirmed by immunoelectron microscopy (IEM) for the presence of Allium viruses. ELISA results showed that the following viruses were detected: Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV) (genus Potyvirus , family Potyviridae ), Garlic common latent virus (GarCLV), Shallot latent virus (SLV) (genus Carlavirus ), Garlic virus D (GarV-D), Garlic virus B (GarV-B) and Garlic virus C type (GarV-C) (genus Allexivirus ). None of the samples reacted with antibodies to Shallot yellow stripe virus (SYSV) genus Potyvirus , family Potyviridae ), Shallot virus X (ShVX) and Garlic virus A (GarV-A, genus Allexivirus ). GarCLV, SLV, GarV-D, GarV-B and GarV-C are reported for the first time from Allium crops in Iran.     

京郊大蒜病毒病的研究及其鳞茎中病毒的脱除

 京郊大蒜病毒病发生普遍,主要症状为条纹花叶、矮化和叶片扭曲畸形。病体细胞中含大量线状病毒粒体和风轮状内含物。线状病毒粒体长度范围为250-1875nm,以长度550-800nm粒体居多。其中长700-800nm的粒体被鉴定为大蒜花叶病毒(GMV),回接脱毒大蒜叶片产生条纹花叶症状。血清学鉴定表明GMV与洋葱黄矮病毒(OYDV)有近缘关系。长度500-600nm的粒体可能为大蒜潜隐病毒(GLV),回接脱毒大蒜不产生花叶症状。下述两种病毒为京郊大蒜的主要病毒。此外,个别标样含烟草花叶病毒,但不是大蒜的主要病毒。对800nm以下线状病毒粒体归属尚待研究。
应用营养茎尖、生殖茎尖和根尖分生组织培养技术,可以脱去上述主要病毒,获得脱毒大蒜。     

番茄斑萎病毒系统侵染我国大蒜

番茄斑萎病毒Tomato spotted wilt tospovirus(TSWV)是严重危害世界经济作物的一种病毒,寄主范围广泛。我们在研究中发现番茄斑萎病毒能侵染我国大蒜Allium sativum L.。利用摩擦接种法将TSWV接种到健康大蒜上,结果显示:接种14 d后大蒜新生叶片出现褪绿和白色斑点症状。ELISA检测显示大蒜叶片汁液与TSWV的单克隆抗体产生血清学反应,采用TSWV N基因的引物对大蒜叶片总RNA进行RT-PCR,结果扩增出约800 bp的条带,在NCBI上BLAST显示与TSWV YN5576的同源性最高,为98.52%。这些数据表明番茄斑萎病毒系统侵染我国大蒜。     

Viral reinfection affecting bulb production in garlic after seven years of cultivation under field conditions

Six sequential field experiments were conducted from 1999 to 2002 to evaluate virus reinfection in garlic cv. Amarante and its effect on bulb production. The treatments in the year 1999 were: T₁ – virus-free garlic-seed obtained by thermotherapy and meristem-tip culture and indexed for virus by immuno-sorbent electron microscopy (ISEM) under its first field cycle; T₂, T₃ and T₄ – garlic-seed in the second, third and fourth field cycles, respectively; and C – standard garlic-seed from the grower (with no control of virus infection) as a control. In the years 2000 to 2002, a new plot of virus-free seed was added to the experiment and cloves from the previous treatments were again grown under field conditions. In the fourth year of experiments, the treatments comprised T₁ (virus-free garlic seeds under the first field cycle) to T₇ (garlic-seed in the sixth field cycle) and C. Two experiments were conducted in the years 1999 and 2000, at two locations and in the years 2001 and 2002 only one experiment per year at one location. The combined analysis of variance for all experiments indicated a significant difference between the treatments for plant height and yield. The bulbs were classified into commercial classes according the Brazilian regulation and the commercial classes 4–7 were 72 %, 60%, 59%, 53% and 35% of the total number of bulbs harvested, from T₁ to T₅, respectively. Virus reinfection at the end of the second field cycle, determined by serology using antisera against the most common viruses of garlic in Brazil reached 83%. Treatments T₁–T₇ yielded 118%, 79%, 57%, 51%, 39%, 33% and 31% higher than the control.     

Double-edged effects of the cryogenic technique for virus eradication and preservation in shallot shoot tips

Plant virus eradication is a prerequisite for the use of virus-free propagules for sustainable crop production. In contrast, virus preservation is required for all types of applied and basic research of viruses. Shoot tip cryopreservation can act as a double-edged strategy, facilitating either virus eradication or virus preservation in cryoderived plants. Here, we tested the efficacies of shoot tip cryopreservation for virus eradication and preservation in shallot (Allium cepa var. aggregatum). In vitro stock shallot shoots infected with onion yellow dwarf virus (OYDV) and shallot latent virus were thermotreated for 0, 2, and 4 weeks at a constant temperature of 36℃ before shoot tip cryopreservation. Results showed that viruses were preserved in recovered shoots when thermotherapy was not applied. Although thermotherapy lowered the regrowth levels of cryotreated shoot tips, the efficiency of virus eradication increased from 5% to 54%. Immunolocalization of OYDV and histological observation of cryotreated shoot tips showed the high frequency of virus preservation was due to the viral invasion of cells close to the apical meristem and the high proportion of cells surviving. Four weeks of thermotherapy drastically decreased the distribution of OYDV, as well as the percentage of surviving cells within the shoot tips, thereby promoting virus eradication. Virus-free plants obtained from combining thermotherapy with cryotherapy showed significantly improved vegetative growth and bulb production. The present study reports how thermotherapy can act as a trigger to facilitate either the safe preservation of Allium viruses or the production of virus-free shallot plants.     

Testing garlic cloves and bulblets for onion yellow dwarf virus by ACP-ELISA

Onion yellow dwarf virus (OYDV) was detected in cloves and aerial bulblets of garlic (Allium sativum) at levels as high as or higher than in leaves of plants grown from tested cloves. It is recommended to test bulblets or a few cloves per bulb before planting to determine if all cloves of a bulb are virus-free. This aids in early detection and allows a more thorough testing of stock than field testing.     

Comparison of Physical, Chemical and Biological Methods of Controlling Garlic White Rot

Treatment of garlic cloves with tebuconazole (at 1ml of Folicur 25% l^–1) achieved a significant reduction in the rate of disease progress and the final incidence of plant death by Sclerotium cepivorum: garlic yields were improved. Although soil solarization provided the best control of garlic white rot, bringing soil populations of S. cepivorum to negligible levels, similar levels of disease control and garlic yields were achieved when tebuconazole was sprayed to stem bases of plants grown from cloves also treated with tebuconazole. This double treatment almost doubled the yield compared with untreated plants and significantly increased bulb quality under high disease pressure conditions. Soil solarization was also highly effective in a second consecutive crop of garlic, with significant improvements in yield and garlic quality. In contrast, lower levels of disease control were obtained when selected isolates of Trichoderma harzianum and Bacillus subtilis were applied to the soil and cloves respectively.     

Survey and characterization of potyviruses and their strains of Allium species

Nearly 5700 plants of 14 cultivated and 8 wildAllium species and varieties from the Netherlands and other parts of the world, were tested for infection with aphid-borne potyviruses by ELISA, electron microscope decoration tests and/or inoculation onto test plants. This resulted in the detection of two known viruses, viz. leek yellow stripe virus (LYSV) and onion yellow dwarf virus (OYDV), and the discovery and characterization of two new viruses, viz. shallot yellow stripe virus (SYSV) and Welsh onion yellow stripe virus (WoYSV), and of six strains of these viruses. ‘Garlic mosaic’, ‘barlic yellow streak’, ‘onion mosaic’, ‘shallot mosaic’, ‘shallot X’, and ‘shallot yellows’ viruses, incompletely described in the literature, are now reidentified as well-known viruses or as strains or mixtures of such viruses. ‘Garlic yellow stripe virus’ is also a complex containing a potyvirus possibly differing from the viruses found in this survey. The symptoms of the potyviruses studied varied widely and ranged from mild to severe chlorotic to yellow striping of leaves, and they are of little diagnostic importance.LYSV was found in vegetatively propagated pearl onion (A. ampeloprasum var.sectivum) from Europe and Asia. It has decreased in leek crops (A. ampeloprasum var.porrum) in the Netherlands since the 1970, apparently due to resistance in new cultivars. OYDV was common in onion (A. cepa var.cepa) from the former USSR and North Africa, and in European cultivars of shallot (A. cepa var.ascalonicum), with the exception of the highly resistant ‘Santé’, but was not detected during this survey in Asian shallot. European samples of ever-ready onion (A. cepa var.perutile), multiplier onion (A. cepa var.aggregatum) and tree onion (A. cepa var.viviparum) contained OYDV. It was also found in sand leek (A. scorodoprasum) from european gene collections. A strain of OYDV from onion and shallot in Morocco and Spain was virulent on onion and shallot cultivars resistant to common OYDV, as reported early for a similar isolate in the USA.Asian shallot appeared generally infected with the new SYSV, similar to OYDV in host range and symptoms but serologically distinct. It was not detected in onion and shallot from Europe or North Africa. A virulent strain of this virus caused striping in sap-inoculated garlic (A. sativum) and Formosan lily (Lilium formosanum). The new WoYSV, infecting Welsh onion in Indonesia and Japan, was earlier described in Japan as OYDV from rakkyo and Welsh onion. It appeared serologically closely related to SYSV and distantly to OYDV, but differed in its host range.Host-specific strains of LYSV and OYDV were detected in garlic, wild garlic (A. longicuspis), an unidentifiedAllium species (suffix-G), and great-headed garlic (A. ampeloprasum var.holmense) (suffix-GhG)., LYSV-G and OYDV-G infected on average 45% and 73%, respectively, of the garlic samples of worldwide origin. Symptoms of isolates of both strains varied in severity, implying the necessity of serological tests for disease diagnosis and health certification. LYSV-GhG was the cause of yellow striping in 93% of the great-headed garlic plants tested, mainly from the Mediterranean area. One sample was also infected with OYDV-GhG.Many samples from vegetatively propagated crops grown from non-certified planting stock contained a few plants free of potyviruses, implying the possibility to obtain healthy (and possibly resistant) selections of such cultivars avoiding meristem-tip culture. Cross-protection of garlic sets by a mild potyvirus isolate seems to be an alternative to the use of vulnerable virus-free sets.Generally, viruses and virus strains could not be transmitted to anyAllium species other than their natural host, except to the highly susceptible crow garlic (A. vineale). This species, and other predominantly vegetatively propagating wildAllium spp. (field garlic,A. oleraceum; ramsons,A. ursinum; sand leek), were found not to be reservoirs of viruses that might infectAllium crops in the netherlands. Streaking in vegetatively propagated wild leeks (A. ampeloprasum and closely related species) originating from the Mediterranean area and Asia was due to an undescribed miteborne virus. The survey confirmed that spread of potyviruses inAllium crops in the Netherlands is from planting sets, and from a neighbouring crop only if of the same species.     

Crude garlic extract significantly inhibits replication of grapevine viruses

Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses.     

甘薯脱毒技术及增产效果研究

作者于１９８８～１９９５年研究了甘薯病毒及甘薯脱毒技术,明确了侵染山东甘薯的主要病毒种类是甘薯羽状斑驳病毒和甘薯潜隐病毒,在国内首次分离侵染甘薯的烟草花叶病毒,探明了其生物学特性。筛选出适合山东甘薯茎尖培养基的最佳激素配比,浓度,ｐＨ值。探索了脱毒薯的增产机理和增产效果,提出了组织培养,茎尖苗检测,脱毒薯速率与推广应用的配套技术规程,培养出徐薯１８等１２个品种的脱毒苗,平均增产４２．９％出干率提高     

Effects of sweet potato feathery mottle virus and sweet potato sunken vein virus on sweet potato yields and rates of reinfection of virus-Free planting material in Israel

Yield reductions ofca 50% or more were observed in field plots infected with both sweet potato feathery mottle virus (SPFMV) and sweet potato sunken vein virus (SPSVV) (‘complex’), compared with plots planted with virus-free propagation stocks. No yield reductions were observed in a plot planted with SPFMV-infected cuttings. In plots infected with SPSVV alone, no significant effect on tuber yields was observed in one year, whereas in the second year there was aca 30% reduction in yield compared with virus-free control plants. Reinfection in the field, in the absence of introduced infection sources, was observed only with SPSVV. However, natural spread resulted when SPFMV-infected source plants were introduced. This implies that aphid vectors were present during the growing season, but that SPFMV infection sources were absent from the area.     

Mite-borne virus isolates from cultivated Allium species,and their classification into two new rymoviruses in the family Potyviridae

While testing several samples of onion and of vegetatively propagated garlic, sand leek and shallot from a number of countries, virus isolates with unusually flexuous particles were obtained by mite (Aceria tulipae) or sap transmissions. No aphid-borne poty-or carlavirus was transmitted by mites, and mite-borne virus isolates could not be transmitted by aphids. The mite-borne isolates did not react with antisera to aphid-borne potyviruses ofAllium spp. or with the Agdia potyvirus group monoclonal. In contrast to the mite-borne onion and garlic mosaic viruses reported in the literature, our mite-borne isolates induced no visible or only very mild symptoms inAllium spp., except isolates from shallot ‘Santé’ which caused diffuse striping. Heavily mite-infested test plants or plant samples showed streaking and malformation due to mite feeding (tangle-top). The mite-borne virus isolates could be classified with test plants and a discriminating antiserum into three groups, representing two viruses and a strain of one of them. They are tentatively named onion mite-borne latent virus (OMbLV), garlic strain of this virus (OMbLV-G), and shallot mite-borne latent virus (SMbLV). Mite transmission, length of virus particles (ca. 700 to 800 nm), and the presence of granular inclusion bodies in infected tissue indicate that the viruses belong to the mite-borne genusRymovirus of the familyPotyviridae. OMbLV from shallot and onion, and OMbLV-G from garlic and sand leek, can be assayed onChenopodium murale but differ in their natural hosts. They are very common. SMbLV, to whichC. murale does not react, was isolated from shallot originating from Asia and Russia.     

Identification and Partial Characterisation of <Emphasis Type="Italic">Lettuce big-vein associated virus</Emphasis> and <Emphasis Type="Italic">Mirafiori lettuce big-vein virus</Emphasis> in Common Weeds Found Amongst Spanish Lettuce Crops and their Role in Lettuce Big-vein Disease Transmission

The potential role of 10 frequently occurring weed species found amongst Spanish lettuce crops as host plants for the two viruses associated with the lettuce big-vein disease, Lettuce big-vein associated virus (LBVaV) and Mirafiori lettuce big-vein virus (MLBVV), was studied. The results showed that both viruses can infect naturally growing Sonchus oleraceus (common sowthistle) plants, the unique susceptible species detected among the analysed weeds. The sequences of the coat protein (CP) genes of the LBVaV and MLBVV isolates recovered from S. oleraceus plants were determined. Phylogenetic studies revealed a very close relationship between the CP sequences from these weed isolates and those from Spanish lettuce. Moreover, we showed that S. oleraceus can act as a source of lettuce infection by means of Olpidium brassicae, the vector fungus of both viruses.     

Efficiency of eradication of four viruses from garlic (Allium sativum) by meristem-tip culture

Mechanical inoculation tests and ELISA with sap from garlic plants used for sanitation by meristem-tip culture revealed four viruses, viz. garlic common latent virus (GCLV) (carlavirus), the garlic strains of leek yellow stripe virus (LYSV-G), onion yellow dwarf virus (OYDV-G) (aphid-borne potyviruses), and onion mite-borne latent virus (OMbLV-G) (taxonomically unassigned virus). The same tests performed on explants grownin vitro showed elimination efficiencies of 100% for LYSV-G, 92% for OYDV-G, 62% for GCLV, and less then 54% for OMbLV-G.Meristem tips excised from garlic cloves and bulbils, 0.15–1.0 mm in size, were tested for regeneration and efficiency of virus elimination after transfer to Murashige and Skoog medium. Successful regeneration into plantlets was obtained with 71% of the meristems from cloves and 72% of those from bulbils, but virus elimination was easiest from cloves: 38% of all explants from cloves and 25% of those from bulbils were virus-free. The efficiency of elimination increased with increasing weight of the cloves, irrespective of the virus. Small tip size seemed to favour virus elimination, but sizes smaller than 0.4 mm led to increasing failure of regeneration.Micropropagation was most successful when cytokinins were omitted from the medium and the garlic shoot was split. Multiplication factors of 3–6 were obtained.     

Production of virus-free carnations by means of meristem culture

The carnations which are normally grown are practically all infected with various viruses, and therefore attempts were made to obtain virus-free plants of a number of varieties by means of heat treatment and meristem culture. Incubators were designed in which the plants could be subjected to a heat treatment at a temperature of 37°C and at a high degree of humidity of the air. The ultimate percentage of virus-free plants in 1960 amounted to an average of 1.3%. This is sufficient to obtain a virus-free clone, but not enough to build up a virus-free assortment within a short time. The factors which may affect the percentage of successful isolations are discussed, viz. the effect of the season, of varietal differences and of differences between the material supplied by different growers. When the virus-free plants are propagated in practice it is essential that the material should be closely inspected and tested at regular intervals. The new material was found to possess most favourable qualities. It is sold with a special certificate.     

New mite-borne virus isolates from rakkyo,shallot and wild leek species

Flexuous viruses were transmitted from rakkyo (Allium chinense) and wild leek species (especiallyA. commutatum) to plants of crow garlic (A. vineale), by transfer of dry bulb mites. By electron microscope decoration tests using three antisera and by inoculations onto test plants, it was concluded that from each of the two natural host species at least two viruses were isolated. The viruses from wild leeks are both pathogenic onAllium spp. and may be of economic importance. Decoration tests on a virus mixture from shallot obtained earlier, revealed another new mite-borne virus in this species. The mite-borne viruses ofAllium spp. appear to be very common; they are largely diverse and their identification remains difficult.