Occurrence and physico-serological properties of potato virus S (PVS) in Iran

1818 collected samples of potato plant showing virus infection symptoms from 85 fields were tested for PVS infection using DAS-ELISA. Average of infection to this virus varied from 0 to 100%. Least infection was belonging to fields with new introduced varieties. On the other hand native and old introduced cultivars showed heavy infection. In field condition, PVS infected plants didn't show very obvious symptoms, so some infected plants may be missed in field sample collecting. The physical properties of 3 isolates, Avaj, Stanboly and Agria No 15 were determined. TIP 55-60 degrees C, DEP 10(-3) and Liv measured 3-4 days. Ouchterlony agar double diffusion test using SDS was useful for virus detection and precipitation lines didn't show any spur between isolates, although isolates differs slightly in symptomatology. SDS-Page and Western blotting methods used successfully for virus detection and determining and measuring viral protein components.     

Viruses infecting sweet potato in Rwanda: occurrence and distribution

A survey of sweet potato virus diseases was conducted in the major sweet potato production areas in low, medium and high altitude zones of Rwanda. A total of 205 symptomatic and 103 asymptomatic samples were collected from 51 sweet potato fields and assayed for Sweet potato feathery mottle virus (SPFMV), Sweet potato chlorotic stunt virus (SPCSV), Sweet potato mild mottle virus (SPMMV), Sweet potato chlorotic fleck virus (SPCFV), Sweet potato latent virus (SwPLV), Sweet potato caulimo‐like virus (SPCaLV) and Cucumber mosaic virus (CMV) using nitrocellulose membrane enzyme‐linked immunosorbent assay. The viruses detected in the samples were SPFMV, SPMMV, SPCSV, SPCFV and SwPLV. Viruses were detected in 83% and 31% of the symptomatic and asymptomatic samples, respectively. SPFMV was detected in 49% of the samples. SPCSV, the second most common virus, was detected in 28% of samples collected from 73% of the fields. About 19% of the samples were tested positive for SPMMV. Thirteen combinations of multiple virus infections were detected in the samples. Viruses were detected in samples from all the fields surveyed, and the frequency of detection was greatest in samples from low altitude zones.     

Time-resolved fluoroimmunoassay of potato virus M with monoclonal antibodies

Mouse monoclonal antibodies (MAbs) specific for potato virus M (PVM) were prepared and the properties of three of them were studied. MAb M4C1 is IgG2b, it binds with high affinity to PVM coat protein, to purified virus preparations and recognises PVM in infected potato leaves and tubers. MAb M6D5 is IgG2a and also reacts with PVM coat protein, purified PVM and with PVM in potato leaf and tuber extracts. In double-antibody sandwich ELISA (DAS ELISA) MAbs M4C1 and M6D5 reacted with all 17 PVM isolates tested. MAb M7 is IgG2b and recognises PVM only in indirect dot ELISA on nitrocellulose filters and viral coat protein on Western blots. MAbs against PVM were used as capture antibodies and europium-labelled MAbs as conjugates in time-resolved fluoroimmunoassay (EuTRFIA). The standard EuTRFIA curve of PVM detection is approximately linear over a range of PVM concentrations from 0.5 ng/ml to 1000 ng/ml. The lowest PVM concentration detectable in EuTRFIA was 0.5 ng/ml and correspondingly 6 ng/ml in DAS ELISA. The use of the europium chelate label allows PVM detection in potato leaf and tuber sap at dilutions greater than 10^--⁴ with very low background fluorescence. EuTRFIA with MAbs, with either one or two incubations is about 10–20 times more sensitive for PVM detection than is DAS ELISA. PVM and PVX, mixed with healthy potato tuber sap, were simultaneously tested in a single sample at concentrations lower than 10 ng/ml by double-label TRFIA using europium-labelled MAbs to PVM and samarium-labelled MAbs to PVX.     

Association of Bactericera cockerelli (Homoptera: Psyllidae) with "zebra chip," a new potato disease in southwestern United States and Mexico

A new defect of potato, Solanum tuberosum L., "zebra chip," so named for the characteristic symptoms that develop in fried chips from infected potato tubers, has recently been documented in several southwestern states of the United States, in Mexico, and in Central America. This defect is causing millions of dollars in losses to both potato producers and processors. Zebra chip plant symptoms resemble those caused by potato purple top and psyllid yellows diseases. Experiments were conducted to elucidate the association between the psyllid Bactericera cockerelli (Sulc) (Homoptera: Psyllidae) and zebra chip by exposing clean potato plants to this insect under greenhouse and field conditions. Potato plants and tubers exhibiting zebra chip symptoms were tested for phytoplasmas by polymerase chain reaction. Potato psyllids collected from infected potato fields also were tested. Results indicated that there was an association between the potato psyllid and zebra chip. Plants exposed to psyllids in the greenhouse and field developed zebra chip. In the greenhouse, 25.8 and 59.2% of tubers exhibited zebra chip symptoms in the raw tubers and fried chips, respectively. In the field, 15 and 57% of tubers showed symptoms in raw tubers and chips, respectively. No zebra chip was observed in tubers from plants that had not been exposed to psyllids, either in the greenhouse or field. No phytoplasmas were detected from potato plants or tubers with zebra chip symptoms, suggesting that these pathogens are not involved in zebra chip. Of the 47 samples of potato psyllids tested, only two tested positive for the Columbia Basin potato purple top phytoplasma.     

Identification and distribution of viruses infecting sweet potato in Kenya

Four hundred and forty-eight symptomatic and 638 asymptomatic samples were collected from sweet potato fields throughout Kenya and analysed serologically using antibodies to Sweet potato feathery mottle virus (SPFMV), Sweet potato chlorotic stunt virus (SPCSV), Sweet potato mild mottle virus (SPMMV), Cucumber mosaic virus (CMV), Sweet potato chlorotic fleck virus (SPCFV), Sweet potato latent virus (SwPLV), Sweet potato caulimo-like virus (SPCaLV), Sweet potato mild speckling virus (SPMSV) and C-6 virus in enzyme-linked immunosorbent assays (ELISA). Only SPFMV, SPMMV, SPCSV, and SPCFV were detected. Ninety-two percent and 25% of the symptomatic and asymptomatic plants respectively tested positive for at least one of these viruses. Virus-infected plants were collected from 89% of the fields. SPFMV was the most common and the most widespread, detected in 74% of the symptomatic plants and 86% of fields surveyed. SPCSV was also very common, being detected in 38% of the symptomatic plants and in 50% of the fields surveyed. SPMMV and SPCFV were detected in only 11% and 3% of the symptomatic plant samples respectively. Eight different combinations of these four viruses were found in individual plants. The combination SPFMV and SPCSV was the most common, observed in 22% of symptomatic plants. Virus combinations were rare in the asymptomatic plants tested. Incidence of virus infection was highest (18%) in Kisii district of Nyanza province and lowest (1%) in Kilifi and Malindi districts of Coast province.     

Stability of RAPD fingerprints in potato: Effect of source tissue and primers

Variations in random amplified polymorphic DNA (RAPD) profiles from leaf, stem, root, and tuber tissues were observed in case of two glasshouse grown potato cultivars using 40 decamer primers suggesting possible danger of cultivar misidentification. Genomic DNA extracted from the above four tissues of four in vitro grown potato cultivars, however, produced more uniform RAPD fingerprints. A significant effect of random primers on fingerprint uniformity was observed in case of both glasshouse and in vitro grown samples. A new concept of stability index for random primers based on homogeneity of RAPD profiles obtained from different tissues of a single plant have been introduced. It is concluded that RAPD analysis of genomic DNA extracted from any tissue of in vitro grown potato plants using 14 selected decamer primers could be used to develop RAPD fingerprints for identification of Indian potato cultivars.     

Factors affecting the detection of potato leafroll virus in potato foliage by enzyme-linked immunosorbent assay

Using antiserum globulins that reacted only weakly with plant materials, potato leafroll virus (PLRV) at 10 ng/ml was detected consistently by enzyme-linked immunosorbent assay (ELISA). The reaction with PLRV particles was slightly impaired in potato leaf extracts that were diluted less than 10^-1 but not at greater dilutions. Antiserum globulins that reacted more strongly with plant materials could be used satisfactorily for coating microtitre plates but were unsuitable for conjugating with enzyme. The detection end-point of PLRV, in leaf sap of potato cv. Cara plants grown from infected tubers in the glasshouse, was about 10^-2 and the virus was reliably detected in extracts of composite samples of one infected and 15 virus-free leaves. PLRV concentration was much less in extracts of roots or stolons than in leaf extracts. The virus was detected in infected leaves of all 27 cultivars tested. PLRV was readily detectable 2 wk before symptoms of secondary infection developed in field-grown plants of cv. Cara and Maris Piper and remained so for at least 5 wk. Its concentration was slightly greater in old than in young leaves and was similar to that in glasshouse-grown plants. In field-grown plants of cv. Maris Piper with primary infection, PLRV was detected in tip leaves 21–42 days after lower leaves were inoculated by aphids; in some shoots it later reached a concentration, in tip leaves, similar to that in leaves with secondary infection. Symptoms of primary infection developed in the young leaves of some infected shoots but were inconspicuous and were not observed until at least a week after PLRV was detected by ELISA.     

Determination of the races of potato virus X and its host rang in Karaj, Damavand and Ardabil, Iran

Potato virus x (PVX) is found commonly in potato-growing areas, worldwide. It is an economically important virus which causes losses in tuber yield of approximate 5 to 15 percent. In a 2 year survey, potato leaf and tuber samples were collected from various fields in Damavand and Karaj. The initial isolations from potato were made by mechanical inoculation first to Gomphrena globosa L. and later to Dartura stramonium L. It was not transmitted by 2 species of Cuscuta but transmitted mechamically. The isolates were inoculated to Nicotiana glutinosa L. in which they were maintained throughout the work. Sap from infected N. glutinosa was ineffective after dilution to 10-6, after 10 minutes at 70 degrees C and after 10 weeks at room temperature. The virus had filamentous and slightly flexuous particles with a normal length of about 490-500 nm and 12 nm width. According to the symptoms, TIP results and serological comparisons, the compared isolates showed no difference and they belong to XN group. In order to estimate disease incidence, 773 tubers from Damavand area were tested and compared with that in Ardabil area. Disease incidence in Damavand ranged from 1.1-20.9 percent and was lower than disease incidence in Ardabil. In 8 genera of collected weeds from fields of potato and tomato samples by using test plants and serological methods, they didn't show existence of the potato virus x.     

马铃薯病毒一步法多重RT-PCR检测技术的构建

根据马铃薯病毒PVX、PVY、PVA、PLRV的CP基因序列设计4对特异性引物,通过对试剂浓度和反应条件进行优化,建立了能够同步检测PVX、PVY、PVA、PLRV的一步法多重RT-PCR检测方法。该方法对PVX、PVY、PVA、PLRV扩增出的靶带大小分别为732、422、132和336 bp,凝胶电泳易辨别区分。病毒RNA最低检测限度为7.8 pg/μL,对PVM、PVS、AMV、TMV及PSTVd的扩增为阴性。研究结果表明,该方法特异、灵敏,比两步法多重RT-PCR检测更加快速、简便,提高了检测效率,降低检测成本,为马铃薯病毒的高效检测提供了有效手段。     

Identification of PVY-N on potato and tobacco in Tehran and Mazandaran provinces

During two growing seasons in years of 2003 and 2004 potato and tobacco of virus infected plants were collected from fields in Tehran (Damavand) and Mazandaran (Behshahr) provinces. Serological methods of TAS-ELISA and DIBA were performed by using PVY antiserum (DSMZ - Plant Virus Collection; Germany) but only PVY was detected. The strain of samples was determined by using MAb of potato virus Y (AS-0403/1; DSMZ; Germany). The molecular weight of the virus coat protein was approximately 34 kDa in SDS-PAGE and Western blotting. Total RNA was extracted for RT-PCR. Immunocapture RT-PCR and RT-PCR products were 974 bp by using specific primers of PVY. IC-RT-PCR has given the best results.     

黑龙江马铃薯Y病毒P1基因的特点北大核心CSCD

【目的】本研究通过对不同PVY分离物基因的测序及分析,从而了解PVY株系的多样性,进而对PVY病毒的分子检测及防治提供重要的资料和参考。【方法】本研究针对黑龙江15个马铃薯Y病毒样品的P1基因进行克隆测序和进化树分析。【结果】经比对分析,样品被分成两组,有10个样品的基因类型高度同源,且相对保守,是本地区的优势群组,无论是与国内其它地区样品比较还是与国外样品比较,其亲缘关系都有一定距离;而另一组中的5个样品的P1基因与本地优势组群有较大差异,且这5个样品间也有一定的差异,并与国内其它地区和国外一些样品的P1基因序列比较接近。通过比对Gen Bank中已上传的序列提供的PVY株系的信息,得知本次试验的P1基因与PVY^(NTN-NW)株系是相似的,且这15个样品与国内其他样品一样都是由PVY^N株系演变而来。【结论】由P1基因分析表明,PVY受环境影响较大,黑龙江10个样品的PVY在长期的进化中产生了具有地方特点的变化,而后来的5个样品说明中国大部分PVY有可能是跟随国外品种资源的引进进入,同时PVY也随国内不同区域间资源交流和种薯调运而传播。     

Application of enzyme-linked immunosorbent assay to the detection of potato leafroll virus in potato tubers

Factors affecting the detection of potato leafroll virus (PLRV) by enzyme-linked immunosorbent assay (ELISA) in tubers of field-grown potato plants with primary or secondary infection were studied. The reactions of extracts of virus-free potato tubers were minimised by pre-incubating the extracts at room temperature and by careful choice of the dilution of enzyme-conjugated globulin. PLRV was reliably detected in tubers produced by secondarily infected plants of all six cultivars tested. PLRV concentration was greater in heel-end than in rose-end vascular tissue of recently harvested tubers but increased in rose-end tissue when tubers stored at 4°C for at least 5 months were placed at 15–24°C for 2 wk. PLRV occurred at greater concentration in tubers from plants of cv. Maris Piper with natural or experimentally induced primary infection than in tubers from secondarily infected plants; again PLRV concentration was greater in heel-end than in rose-end vascular tissue. Plants whose shoots were infected earliest in the growing season were invaded systemically and produced the greatest proportion of infected tubers; plants infected late in the season also produced infected tubers but PLRV was not detected in their shoot tops. PLRV concentration in tubers from the earliest-infected plants was less than in tubers from later-infected plants. PLRV was detected reliably by ELISA in tubers from progenies that were totally infected but was not detected in all infected tubers from partially infected progenies. ELISA is suitable as a routine method of indexing tubers for PLRV, although the virus will not be detected in all infected tubers produced by plants to which it is transmitted late in the growing season.     

Field trials using single potato plants as whole plots, with and without nematicide treatment, to assess tolerance of attack by Globodera rostochiensis

It is important to assess the tolerance of new potato cultivars with partial resistance to potato cyst nematode attack. The experiments described here are intended to quantify tolerance adequately using a minimum of resources. When test plants were spaced widely in the field trials it was also possible to estimate their resistance levels but standard plant spacing allowed roots of neighbouring plants to encroach on a test plant's root space and to interfere with attempts to assay resistance. Standard plant spacing did, however, give more realistic yields per plant and competition from neighbouring plants was standardised by surrounding each test plant by four identical guard plants (cv. Désirée). Regressions of yield on severity of nematode attack were drawn when nematode infestation varied naturally across the trial site or when different levels of attack were created by applying nematicide at a range of dose rates. Cara was the most tolerant cultivar tested, with a complete spectrum of tolerance shown by other cultivars/clones. Some clones with a high degree of resistance were intolerant and it is precisely these that this type of test seeks to identify.     

A glasshouse test to assess the sensitivity of potato cultivars to tobacco rattle virus

The sensitivity to tobacco rattle virus of nine potato cultivars and 24 potato clones from the potato breeding programme at SCRI was assessed in a glasshouse pot test and in field trials for two consecutive years. The results and analyses presented indicate that the pot test gives an accurate and reliable estimate of sensitivity to tobacco rattle virus. Methods of assessing severity of expression of disease symptoms were examined. The advantages of such a glasshouse pot test and its application are discussed.     

Local variation in susceptibility of Colorado potato beetle (Coleoptera: Chrysomelidae) to insecticide

The susceptibility status of Colorado potato beetle, Leptinotarsa decemlineata (Say), adults to phosalone was determined by dip and glass jar assay techniques. Bioassay results indicated a narrow variation in Colorado potato beetle insecticide susceptibility among sample sites. LC50 values were generally highest from specimens collected in field that received frequent phosalone applications for seven consecutive growing seasons. In five populations tested, LC50 values ranged from 503.72 to 827.95 ppm in dip test method. In glass jar technique, resistance ratio value of 1.72 for LC50 was obtained. A significant linear relationship between LC50 values of individual populations across test methods was detected. Both bioassay techniques were suitable for monitoring resistance to insecticide in Colorado potato beetle adult populations. Glass jar technique, however, exhibited less variability in LC50 estimates and showed a higher degree of sensitivity than the dip method. Filter paper and leaf disk techniques for larvae were two bioassay methods used to determine phosalone susceptibility in L. decemlineata populations. Both bioassay techniques exhibited a similar level of susceptibility of the larvae to phosalone; however, the fiducial limit values from filter paper method were narrow than the leaf disk assay technique. A significant direct relationship between LC50 values of individual population across test methods was observed. Differences in LC50 ranking among fields between adults and larvae indicated a differential susceptibility to insecticide between life stages. Low LC50 values obtained from Colorado potato beetle in sample sites indicated that phosalone resistance was not severe in these fields. The glass jar and filter paper testing methods are simple and sensitive test techniques for measuring susceptibility of Colorado potato beetle adults and larvae to phosalone, respectively.     

Genetically engineered resistance to potato virus X in four commercial potato cultivars

The genes for the capsid protein (CP) and the 8K movement protein of PVX were introduced into potato (Solanum tuberosum L.) and expressed under the control of CaMV 35S promoter using a binary vector andAgrobacterium tumefaciens. Four commercial potato cultivars (Russet Burbank, Shepody, Desirée and Bintje) have been efficiently transformed. Eleven independent transgenic clones, with CP expression levels higher than 0.05% of the soluble leaf proteins, were analyzed for resistance to inoculation with PVX (5 and 50µg/ml). The resistance of the transgenic plants to PVX was observed with the lower titer of virus inoculation (5 µg/ml) but not with higher titer (50 µg/ml). A significant reduction in the accumulation of virus in the inoculated transgenic potato plants has been observed under greenhouse and field conditions. Furthermore, the CP gene is very stable and is transferred to new plants originated from stem cuttings or from tubers. The transgenic plants appeared to be phenotypically identical to the nontransformed controls.Abbreviations BAP benzyl-aminopurine - BCIP 5-bromo-4-chloro-3-indolylphosphate p-Toluidine salt - CaMV cauliflower mosaic virus - CP capsid protein - GA₃ gibberellic acid - Kbp kilobase pair - NAA naphthalene acetic acid - NBT nitroblue tetrazolium chloride - NOS nopaline synthase - NPT II neomycin phosphotransferase II - PMSF phenyl methyl sulfonyl fluoride - PVX potato virus X - PVY potato virus Y     

Occurrence and distribution of viruses infecting potato in Russia

Potato viral disease has been a major problem in potato production worldwide including Russia. Here, we detected Potato Virus M (PVM), P (PVP), S (PVS), Y (PVY), and X (PVX) and Potato Leaf Roll Virus (PLRV) by RT-PCR on potato leaves and tubers from the Northwestern (NW), Volga (VF), and Far Eastern (FE) federal districts of Russia. Each sample was co-infected with up to five viruses. RT-PCR disclosed all six viruses in NW, three in VF, and five in FE. Phylogenetic analyses of PVM and PVS strains resolved all PVM isolates in Group O (ordinary) and all PVS isolates in Group O. Seven PVY strains were detected, and they included only recombinants. PVY recombinants were thus the dominant potato virus strains in Russia, although they widely varied among the regions. Our research provides insights into the geographical distribution and genetic variability of potato viruses in Russia.     

Detection,distribution and control of Potato mop-top virus,a soil-borne virus,in northern Europe

Potato mop-top virus (PMTV; genus Pomovirus; family Virgaviridae) is transmitted by the soil-borne Spongospora subterranea f.sp. subterranea, a protoctist that causes powdery scab on potato. PMTV is distributed widely in the potato growing areas in South and North America, Japan and northwestern Europe. This article reviews the current knowledge on detection, distribution and control of PMTV with focus on the Baltic Sea region. Since the 1980s, PMTV has caused great economic losses to potato production in the Nordic countries (Norway, Sweden, Denmark and Finland), but its occurrence in other countries of the Baltic Sea region remained unknown. To fill this knowledge gap, harmonised sampling and virus detection procedures including bioassays and serological and molecular methods were employed by 21 research institutions to detect PMTV in potato tubers and soil samples in 2005–2008. Potato growing areas were widely contaminated with PMTV in the Nordic countries. Only the main seed potato production area in northern Sweden and the High Grade seed potato production zone in Finland were negative for PMTV. Intensive and systematic surveys in Poland in 2004–2008 found no evidence of PMTV, except a single PMTV-infected tuber detected in 2008. Surveys in the Baltic countries (Lithuania, Latvia and Estonia) and northwestern Russia (Leningrad province) were negative for PMTV, except infection of minitubers in a screenhouse in Latvia in 2005. Varying percentages of tubers expressing spraing symptoms in Sweden, Norway, Denmark and Poland were infected with Tobacco rattle virus, and bioassays indicated similar results for Russia. Incidence of symptomless infections with PMTV was high in tubers of many potato cultivars. Here, we discuss the contrasting patterns of distribution of PMTV in the Baltic Sea region, factors playing a role in dispersal and establishment of PMTV in new fields and means for controlling PMTV and its spread to new areas. We emphasise the use of the current virus-specific methods for the detection of PMTV in symptomless potato tubers and the high risks of disseminating PMTV to new fields and areas in viruliferous resting spores of S. subterranea in the soil adhering to seed tubers. PMTV-resistant potato cultivars will provide the only sustainable means for preventing yield losses in the infested fields and the prospects of resistance breeding are summarised.     

Aspects of resistance to sweet potato virus disease in sweet potato

In field trials during the first and the second rainy season of 1996 in Uganda, whiteflies were similarly abundant and aphids were absent on three clones of sweet potato (NIS-93–63, cv. Tanzania and cv. New Kawogo) although the three clones differed considerably in their resistance to sweet potato virus disease (SPVD), a complex disease resulting from infection by both the aphid-borne sweet potato feathery mottle virus (SPFMV) and the whitefly-borne sweet potato chlorotic stunt virus (SPCSV). This suggests that vector resistance does not determine the relative SPVD resistance of these genotypes. SPFMV alone had only a low virus titre in sweet potato cvs Tanzania and New Kawogo, became increasingly difficult to detect in plants of these cultivars and was seldom acquired by aphids. However, this resistance to SPFMV was not apparent in plants which were also infected with SPCSV. Plants then had a high SPFMV titre, appeared unable to eliminate SPFMV and provided good sources for aphids to acquire it.     

Control of potato cyst nematode (Globodera rostochiensis) by nematicides and a resistant potato cultivar at four sites in Scotland

The control of potato cyst nematode (Globodera rostochiensis) by the oxime-carbamates aldicarb and oxamyl was tested in four fields in Scotland. Dazomet was tested in three of these fields and carbofuran in one. In untreated plots in the three most heavily infested fields Maris Piper (resistant) yielded better than Pentland Crown (non-resistant). All nematicides increased the yields of both potato cultivars but had a greater effect on the yield of Pentland Crown. Dazomet increased yields of tubers most. Heavy nematode infestation reduced yield of tubers more in a sandy soil than in two sandy loams. In a field with few potato cyst nematodes nematicides did not significantly affect tuber yields. Although the nematicides greatly increased yields, they were not completely effective in controlling potato cyst nematodes. In treated plots in the lightly infested field, there were more nematode eggs following a crop of Pentland Crown than before. In contrast, Maris Piper markedly decreased post-cropping populations and except at one site, where dazomet further decreased nematode numbers, combining nematicides with the resistant cultivar failed to decrease nematode numbers further. Nematicides decreased the numbers of larvae invading potato roots by up to 95%, oxamyl at 5–6 kg/ha being consistently the best treatment.