Dissemination and detection of peanut clump virus in groundnut seed

Experiments were done to assess the role of seed-transmission in the dissemination of peanut clump virus (PCV) in groundnut (Arachis hypogea L.), and the usefulness of enzyme-linked immunosorbent assay (ELISA) for detecting the virus in infected groundnut seed. The virus was present in 7.5% of seedling progeny from infected plants and could be detected in 16.5% of the seeds by ELISA. When groundnut seeds were grown in a field contaminated by the virus, it was shown that by roguing the infected plants, only 0.1% of the seeds from the remaining plants contained the virus. It was also established that the level of contamination of seeds by the virus was inversely proportional to the seed size.     

Rapid detection of cowpea aphid-borne mosaic virus in cowpea seeds

Four cultivars of cowpea (Vigna unguiculata [L]. Walp.) were infected with cowpea aphid-borne mosaic virus (CABMV) by natural infection in field plots. Seeds taken from these plants were tested for the presence of the virus by ELISA and symptom observation on the plantlets grown from the seeds. A biotin/ streptavidin ELISA technique was used and found to be more sensitive than a standard ELISA protocol for detecting CABMV infection in seed. There was a good correlation between the ELISA detection of CABMV in tissue taken from single cowpea seeds and subsequent development of infected plants grown from the same seeds. The ELISA technique is reliable for selecting CABMV-free stocks of cowpea seeds.     

Occurrence of Melon Necrotic Spot Virus in Crete (Greece)

Since 1982 melon plants cv. Galia grown in plastic houses showed severe leaf and stem necrosis. A virus isolated from affected plants infected only Cucurbitaceae. Systemic infections were only developed in melon and Luffa acutangula, while a severe hypocotyl necrosis was observed in watermelon. Purified virus sedimented in sucrose gradients as a single component and reacted only with antisera to melon necrotic spot virus (MNSV). Seed transmission (22.5%) of the virus was observed m melon plants grown from seed of naturally infected melons, but the virus could not be detected in triturated seeds. The virus could be isolated from leachate of contaminated soil and melon plants became infected when grown in contaminated soil or when watered with suspensions of virus. These properties suggest that the virus is an isolate of MNSV.     

The Association of Brome Mosaic Virus and Wheat Rusts: I. Transmission of BMV by Uredospores of Wheat Stem and Leaf Rust

Both the stem and leaf rust fungi of wheat (Puccinia graminis f. sp. tritici, P. recondita f. sp. tritici) were investigated as potential vectors of brome mosaic virus (BMV), a pathogen of Gramineae. They were found to transmit BMV to healthy plants through uredospore transfer from hosts infected with both the viral and fungal pathogens. Virus was transmitted even to rust-resistant and non-susceptible hosts. Rust-transmitted virus produced atypical symptoms on plants, could not be extracted by normal low pH methods, and multiplied only to low levels in leaf tissue. Extracted virus was shown to be similar to BMV-type strain (ATC) by serology, density gradient centrifugation and immunospecific electron microscopy. The importance of the association of BMV with the rust fungus in natural infections is discussed.     

Response of wheatgrasses and wheat × wheatgrass hybrids to barley yellow dwarf virus

Summary Resistance to barley yellow dwarf virus (BYDV), manifested by low enzyme-linked immunosorbent assay (ELISA) values in plants exposed to viruliferous aphids, was identified in several wheatgrasses (Agropyron spp.). ELISA results were similar for root and leaf extracts of infested plants. No difference in reaction to BYDV was found between plants grown in the field and those in the growth chamber. Interspecific hybrids were generated using pollen from single resistant plants of Agropyron spp. to pollinate soft red winter wheat spikes. Resistance in hybrids appeared to be at the level of virus replication rather than at the level of vector inoculation. The hybrids varied in their reaction to BYDV. Expression of BYDV resistance in hybrids was influenced not only by wheat genotype and Agropyron species but, in some cases, reaction varied even among hybrids between the same wheat genotype and Agropyron plant. Implications of these results are discussed.Contribution from the Purdue Univ. Agric. Exp. Stn., West Lafayette, IN 47907, and USDA-ARS. The research was supported in part by Public Varieties of Indiana. Purdue Univ. Agric. Exp. Stn. Journal No. 11656     

Use of Clq enzyme-linked immunosorbent assay to detect plant viruses and their serologically different strains

Clq was prepared from bovine serum using a simple method involving repeated dialysis at low ionic strength in the presence of chelating agents (yield c. 3 mg/100 ml serum). It was viable when stored at -18°C for up to 2 months, and at 4°C for at least 10 wk in a storage buffer containing 10% sucrose. When used in Clq ELISA this test was as sensitive as the direct double antibody sandwich form of ELISA (direct ELISA) in detecting purified potato virus Y (PVY), with a limit of detection in both methods of c. 15 ng/ml, and slightly more sensitive in detecting purified cocksfoot mild mosaic virus (CMMV), with limits of detection of c. 15 ng/ml and c. 15–60 ng/ml respectively. Using an antiserum to one strain of each virus, Clq ELISA readily detected strains of PVY, CMMV, Andean potato latent virus (APLV) and barley yellow dwarf virus (BYDV). This included detection of APLV-Hu by APLV-Caj antibodies and CMMV(G) by PMV(S) antibodies, neither of which system gives detection in direct ELISA. Clq ELISA was therefore less specific than direct ELISA in detecting serologically different virus strains. Virus detection by Clq ELISA was inhibited when sap of tobacco, Nicotiana clevelandii and Setaria italica was used at low dilution. Inhibition by N. clevelandii sap was alleviated by using increased concentrations of virus specific antibody to detect APLV and plum pox virus. Also, extracting APLV infective N. clevelandii or CMMV infective S. italica saps in a minimum of buffer, centrifuging at low speed and diluting the supernatant before testing, partially overcame the inhibition. The inhibitory substance(s) in sap may act by preventing the binding of Clq to virus-antibody aggregates. Sap of wheat, oat and barley did not appear to have an inhibitory effect and BYDV was readily detected in naturally infected field grown plants of these species.     

Tests for transmission of four potato viruses through potato true seed

The Andean potato calico strain of tobacco ringspot virus (TRSV-Ca) was detected in 2–9% of potato seedlings grown from true seed from plants of cv. Cara and clone G5998(6) infected with TRSV-Ca. Similarly, a potato isolate of the oca strain of arracacha virus B (AVB-O) was detected in 4–12% of progeny seedlings of cv. Cara and clone D42/8 infected with AVB-O. Potato virus T (PVT) passed through 33–59% of seed from PVT-infected cv. Cara, but only 0–2% infection was detected in seedlings from seed of PVT-infected clone D42/8. By contrast, no infection was detected in seedlings grown from seed from plants of G5998(6), D42/8 or cv. Cara infected with Andean potato latent virus strains Hu (APLV-Hu) or Caj (APLV-Caj), although both strains passed through seed of Nicotiana clevelandii. AVB-O, PVT and TRSV-Ca were detected in all tests of pollen from flowers of infected potato plants, but APLV-Hu and APLV-Caj were detected less frequently. AVB-O and PVT were transmitted through 2% and 8% respectively, of seed from healthy potato plants pollinated with pollen from infected plants. However, no transmission through seed was detected when pollen from TRSV-Ca infected plants was used. None of the four viruses were transmitted to healthy potato plants pollinated with pollen from infected plants. APLV-Hu caused exceptionally severe symptoms in the cv. Cara plants used for seed production, but the Bolivian strain of PVT induced only mild symptoms rather than the severe systemic necrosis previously reported for the type of strain of PVT in this cultivar. No symptoms developed in potato seedlings infected with TRSV-Ca, AVB-O or PVT through the seed.     

Seed transmission of turnip yellow mosaic virus in winter turnip and winter oilseed rapes

This is the first record of seed transmission of turnip yellow mosaic virus (TYMV) in oilseed and turnip rapes. The seed transmission of TYMV in a naturally infected winter turnip rape (Brassica napus var. silvestris) cultivar Perko PVH was investigated. By ELISA 1.6%, 3.2% and 8.3% seed transmission of the virus was found in seed of plants from three localities. The proportion of infected seeds produced by artificially infected plants of winter oilseed rape (Brassica napus ssp. oleifera) and winter turnip rape cultivars was determined. The virus transmission rate, expressed as the proportion of virus-infected plants which germinated from the seed was for the oilseed rape cvs Jet Neuf 0.1%, Solida 0.4%, Silesia 0.8%, Darmor 1.2%, SL-507 0.2%, SL-509 0.0% and for the winter turnip rape cv. Perko 1.5%. ELISA cannot be used in direct tests on bulk seed lots to estimate proportion of infected seed, but must be used on germinated seedlings.     

The uptake of phorate, a systemic insecticide, applied as a slurry to wheat and mustard seeds

The absorption by plants of wheat and mustard of a systemic organophosphorus insecticide (phorate), applied as a slurry seed dressing, was studied by caging the aphid Rhopalosiphum padi (L.) on the foliage of wheat, and the aphid Brevicoryne brassicae (L.) and the Chrysomelid beetle Phaedon cochleariae (F.) on white mustard, grown from phorate-treated seed.
Wheat and mustard plants quickly lost their toxicity to insects when they were transplanted, suggesting that most of the insecticide from a slurry seed treatment passes into the soil and is picked up by the roots. That phorate or its derivatives occur in the soil was shown by tests of anticholinesterase activity. Insecticide can also pass into the seed of wheat and move to the growing embryo. Phorate becomes closely bound to the testa of mustard, but does not penetrate it to reach the cotyledons or other parts of the embryo. Mustard cotyledons can become contaminated by insecticide as they emerge through the soil.
Young and old leaves of both wheat and mustard depend on continued absorption of insecticide from the soil to maintain their toxicity. No insecticide moves from old to young leaves. Old leaves lose their toxicity to insects more slowly than young ones. When treated seeds are sown close together, the overlapping zones of insecticide round each seed can increase strength and persistence of insecticidal effect. This happens more with dimethoate, which readily dissolves in water, than with phorate, which is almost insoluble. At normal sowing rates the zones of insecticide round each seed would rarely overlap.
Roots of wheat and mustard from treated seed did not excrete insecticide, and the roots did not carry insecticide through the soil.     

Survey of two beet viruses in South Kazakhstan and Central Asia

During two surveys of beet root crops in South Kazakhstan and Central Asia conducted in 1988 and 1989, 465 of 990 samples were found to contain beet mosaic virus (BMV) by double-antibody sandwich (DAS) ELISA. BMV infection was widely scattered in the area surveyed, and its incidence varied considerably, reaching 100% in some fields adjacent to beet seed crops. BMV isolates from Kazakhstan, Kirgizia, Uzbekistan and Ukraine were found to be serologically closely related in DAS-ELISA test. Beet yellows virus (BYV) was not detected in any location surveyed in South Kazakhstan and Central Asia. BYV spread into the area is probably prevented by its geographical isolation.     

Preliminary studies on sap-transmissible viruses of red clover (Trifolium pratense L.) in England and Wales

Red clover plants, collected from nine widely separated permanent pastures in England and Wales, were tested for sap-transmissible viruses. Viruses were identified by the symptoms they caused in test plants, by electron microscopy, and by serological tests. Of the 265 plants tested 14% were infected. Only pea mosaic virus was common and widespread; it was found in 8% of the plants, and in seven of the fields. Other viruses isolated were arabis mosaic, bean yellow mosaic, red clover mottle, and red clover vein mosaic; only red clover mottle virus produced diagnostic symptoms in red clover. No viruses were detected in seedlings grown from seed from eighty-nine commercial seed crops. Attempts to transmit red clover mottle virus by the Collembolan Sminthurus viridis L., which is common on red clover, failed.     

Seed phosphorus (P) content affects growth,and P uptake of wheat plants and their association with arbuscular mycorrhizal (AM) fungi

Two experiments were carried out in a growth chamber and a naturally lit glasshouse to investigate the influence of seed phosphorus (P) reserves on growth and P uptake by wheat plants (Triticum aestivum cv Krichauff), and their association with arbuscular mycorrhizal (AM) fungi. Increased seed P reserves improved plant growth at a range of P supply up to over 100 mg P kg^–1 soil. Plants grown from seeds with high P reserves tended to accumulate more P from soil, which was mainly attributed to better root system development. Mycorrhizal colonisation did not significantly affect P uptake of plants grown with low irradiance (in growth chamber). However, in the naturally lit glasshouse, mycorrhizal plants had significantly higher P concentrations than non-mycorrhizal plants. Furthermore, mycorrhizal plants grown from seeds low in P accumulated similar amounts of P compared with those grown from seeds with high P, indicating that mycorrhizal colonisation may overcome the disadvantage of having low seed P reserves in the field.     

Comparison of ELISA and immunoblotting techniques for the detection of cherry mottle leaf virus

Formaldehyde treated cherry mottle leaf virus (ChMLV) and the isolated coat protein were used successfully for the production of polyclonal and monoclonal antibodies. The monoclonal antibodies had a titre of 1:51 200 and consisted of IgG1 and IgG2. The antibodies reacted with all 11 isolates of ChMLV, from five locations in Canada and the USA, included in this study. Several serological procedures were assessed to compare their sensitivity for detecting ChMLV. Plate-trapped antigen ELISA (PTA-ELISA) and dot-blot immunobinding assay (DBIA), using virus specific MAbs, were the most sensitive tests in this study. Triple antibody sandwich ELISA (TAS-ELISA) and Western blot were found to be less sensitive. Dilution of the samples appeared to increase the sensitivity of both PTA-ELISA and Western blot detection. Young leaves and flowers of Prunus avium were the best tissue for detecting the virus which could also be detected in the fruit and leaves of P. tomentosa. April and May were optimal for detection of the virus in the field, whereas both April to May and August to September were optimal for screenhouse-grown plants.     

Further studies on seed transmission of broad bean stain virus and Echtes Ackerbohnenmosaik -Virus in field beans (Vicia faba)

Broad bean stain virus (BBSV) and Echtes Ackerbohnenmosaik-Virus (EAMV) were detected in the seed coat and embryo sac fluid of immature seeds from infected field beans (Viciafaba minor) by inoculation to Phaseolus vulgaris; BBSV was also detected in immature embryos. The proportion of seeds infected with either virus decreased during maturation. The viruses were transmitted to seedlings as often through fully ripened seeds from which the seed coats had been removed as through intact seeds. Both viruses were detected in pollen from infected plants, but in glasshouse tests only BBSV was transmitted through pollen to seeds. Delaying fertilization in plants infected with BBSV or EAMV seemed not to affect seed transmission of either virus. In glasshouse tests BBSV was transmitted more often through seeds from plants that were inoculated before flowering than during flowering, and was not transmitted through seeds from plants inoculated after flowering; EAMV was transmitted only through seeds from plants inoculated before flowering. In tests on seed from naturally infected plants BBSV was transmitted more often through seeds from plants that developed symptoms before flowering than during flowering. Both viruses were seed-borne in all cultivars tested and there was no marked difference in the frequency of transmission of either virus among the spring-sown cultivars most common in Britain. Both viruses persisted in seed for more than 4 yr.     

Studies on the seed transmission of cucumber mosaic virus in chickweed (Stellaria media) in relation to the ecology of the virus

Cucumber mosaic virus (CMV) was transmitted in the seed of infected Stellaria media plants. The rate of seed transmission varied both in manually infected plants (3–21%) and in plants grown from infected seed (21–40%). In naturally infected plants the rates of transmission found were 4–29%. Seeds recovered from field soil carried 4–5% infection and in infected seed placed in the soil the virus persisted for at least 5 months. Seed transmission of CMV also occurred in infected Lamium purpureum (4%), Cerastium holosteoides (2%) and Spergula arvensis (2%) but it could not be demonstrated in six other more common weed species in five botanical families. Seed transmission in Stellaria media occurred with a British (W) and an American (Y) strain of CMV. The virus was shown to occur in S. media pollen. The importance of CMV-infected S. media seed in the soil in relation to the epidemiology of the virus is discussed.     

两种镉积累型小麦根际微生物群落结构及功能多样性

为探究不同积累型小麦品种对根际微生物群落结构及功能多样性的影响,以镉低积累型小麦济麦22和镉高积累型小麦冀5265为研究材料,采用分离培养法和Biolog-Eco微平板法分析根际细菌数量、可培养优势群落结构以及微生物群落功能多样性。结果表明：污染土壤济麦22根际总细菌数量和抗Cd细菌数量均显著高于冀5265,而非污染土壤中两品种间无差异。污染土济麦22根际发现较多产脲酶和高镉抗性菌株(200 mg/L)。污染土济麦22根际优势菌多为Arthrobacter sp.和Bacillus sp.,冀5265根际优势菌主要为Streptomyces sp.;非污染土济麦22与冀5265根际优势菌群相似,均以Bacillus sp.为主。Biolog试验结果表明,两个小麦品种根际微生物群落对碳源的利用能力存在差异,济麦22根际微生物AWCD值、Mc Intosh指数、Shannon-Wiener指数、Simpson指数在污染土和无污染土中均显著高于冀5265。因此,污染土壤中不同积累型小麦品种根际微生物群落结构及功能多样性均存在差异,该研究结果对于揭示高低积累型小麦根际微生物机制提供了重要参考依...     

Viral twig necrosis of sweet cherry. Modes of transmission and spread of petunia asteroid mosaic virus (PeAMV)

Grafting symptomless scions, derived from petunia asteroid mosaic virus (PeAMV)-infected trees, to healthy rootstocks resulted in only 3.3% infection in the resulting trees. Up to 90% of seeds from infected sweet cherries contained high quantities of PeAMV, but the virus was not transmitted to the seedlings apparently because of low virus content in the embryo and loss of infectivity during seed maturation and storage. Replanting healthy cherry trees cv. Sam, grafted to different rootstocks, into contaminated soils resulted in new infections. Eight of 13 trees on rootstocks derived from Prunus avium (F 12/1 and cv. Sam on its own roots) were infected with PeAMV within a period of four years but only one of 16 trees on Weiroot-rootstocks (selections from Prunus cerasus) became infected. The detection of PeAMV in naturally contaminated soil samples by the bait plant procedure, using Nicotiana clevelandii, was superior to testing soil eluates by enzyme-linked immunosorbent assay (ELISA) and immuno electron microscopy (IEM). Wild plants may contribute to virus propagation and maintenance of virus contamination of the soil as 25 of 310 samples from 712 herbaceous plants growing in the vicinity of infected trees contained PeAMV; the contaminated samples represented 12 species. The perpetuation of PeAMV by infected scion wood is probably of minor significance, and infection via the soil probably represents the most important means of spread of viral twig necrosis in northern Bavaria.     

Studies on the occurrence of tomato bushy stunt virus in English rivers

Tomato bushy stunt virus (TBSV) of unknown source was isolated from water of the River Thames, near Oxford. The isolate designated TBSV-T was mechanically transmissible to several tomato (Lycopersicon esculentum) cvs and to other species including Petunia hybrida, pepper (Capsicum annuum). eggplant (Solanum melongena), Nicotiana clevelandii, Chenopodium amaranticolor and C. quinoa in which it caused systemic symptoms. It caused no infection of globe artichoke (Cynara scolymus) or Pelargonium domesticum. The virus was not adsorbed to soil and could be isolated from leachate of soil in which systemically-infected tomato or C. quinoa plants were grown. Tomato plants became infected when grown in soil watered with virus suspensions. TBSV-T was infective after 10 min at 80°C but not at 90°C and when diluted to 10^-5 but not to 10^-6. Purified virus preparations contained C. 30 nm isometric particles. In gel-diffusion serological tests, TBSV-T reacted with homologous anti-serum and with antiserum to petunia asteroid mosaic virus but not to pelargonium leaf curl virus. Seed-borne infection (50–65%) of TBSV was demonstrated in plants grown from seed of symptomlessly-infected tomato fruit. TBSV was isolated from symptomlessly-infected tomato fruit imported from Morocco during October-April 1981. One of the isolates (TBSV-M) was indistinguishable from TBSV-T in host range, symptomatology and serological reactions. TBSV was also found in tomato plants growing extraneously in primary settlement beds at sewage works; such plants having been derived from undigested seeds in sewage. Because of its ‘alimentary-resistance’ in man, it is possible that one ecological route whereby TBSV enters rivers is by man's consumption of TBSV-infected tomatoes and eventual sewage dispersal into rivers.