Observations and experiments on the use of an avirulent mutant strain of tobacco mosaic virus as a means of controlling tomato mosaic

In 1973 tobacco mosaic virus (TMV) strain M II-16 was successfully used by growers in the United Kingdom to protect commercial tomato crops against the severe effects of naturally occurring strains of TMV. However, plants in many crops had mosaic leaf symptoms which were occasionally severe, so possible reasons for symptom appearance were examined. The concentration of the mutant strain in commercially produced inocula (assessed by infectivity and spectrophotometry) ranged from 28 to 1220 μg virus/ml; nevertheless all samples contained sufficient virus to infect a high percentage of inoculated tomato seedlings. Increasing the distance between the plants and the spray gun used for inoculation from 5 to 15 cm resulted in a significant decrease in the number of tomato seedlings infected. When M II-16 infected tomato plants were subsequently inoculated with each of fifty-three different isolates of TMV, none showed severe symptoms of the challenging isolates within 4 wk, although some isolates of strain o induced atypically mild leaf symptoms. In a further experiment, M II-16 infected plants showed conspicuous leaf symptoms only 7 wk after inoculation with a virulent TMV isolate. M II-16 multiplied more slowly in tomato plants and had a lower specific infectivity than a naturally occurring strain of TMV. More than 50% of plants in crops inoculated with strain M II-16 which subsequently showed conspicuous leaf mosaic contained TMV strain 1 or a form intermediate between strains o and 1. It is suggested that the production of TMV symptoms in commercial crops previously inoculated with strain M II-16 may result from an initially low level of infection, due to inefficient inoculation, which allows subsequent infection of unprotected plants by virulent strains. Incomplete protection by strain M II-16 against all naturally occurring strains may also be an important factor.     

Interactions between a seed-borne strain of cucumber mosaic cucumovirus and its lupin host

The relationship between time of inoculation with cucumber mosaic cucumovirus (CMV) and the growth, seed production and rate of seed transmission of virus in lupin (Lupinus angustifolius cv. Illyarrie) was studied in field-grown plants. Plants inoculated at the seedling stage (2 days post-emergence) showed 45% mortality. Plants infected through the seed were more stunted than plants inoculated at the seedling stage. Plants inoculated up to the mid-vegetative growth stage (58 days post-emergence) yielded ≤ 27% of the dry matter and ≤ 9% of the seed of healthy plants. Late inoculation (114 days post-emergence) did not affect dry matter yield, but reduced seed yield to 75% of that of healthy plants. Rate of seed transmission depended on the time of inoculation of plants. The maximum rate was 24.5% for plants that were inoculated at the mid-vegetative growth stage (58 days post-emergence). However, early inoculation caused a large reduction in seed yield, and it was shown that plants inoculated at the beginning of flowering (94 days post-emergence) produced greater numbers of infected progeny than plants inoculated at earlier or later times. No relationship was observed between seed weight and transmission of CMV. Infectious CMV was recovered from the embryo, but not from the testa. A simple seed transmission model was used to evaluate several hypothetical epidemics and to determine the time of inoculation which results in greatest rates of seed transmission of CMV. For example, when fewer than 73% of plants in a crop become infected with CMV, then the rate of transmission of virus in crop seeds will be greatest when inoculations are at the beginning of flowering.     

The effect on yield in courgette and marrow of the mild strain of zucchini yellow mosaic virus used for cross-protection

The effects on yield in courgette and marrow (Cucurbita pepo) crops resulting from inoculation with the mild strain of zucchini yellow mosaic virus (ZYMV:WK), have been determined in polythene-house trials and in three years of outdoor, commercial field trials. In polythene-house trials ZYMV:WK inoculated plants were up to 10 days later in flowering than uninoculated plants and their cumulative yields were between 5% and 26% less than uninoculated plants depending on the cultivar. In most field trials cumulative yields from inoculated plants were between 4% and 38% less than uninoculated plants depending on the site and cultivar, but in one trial the yield was 7% higher from inoculated plants. In all experiments, courgette and marrow fruits harvested from ZYMV:WK inoculated plants were symptomless and indistinguishable from fruit harvested from uninoculated plants. The mild leaf symptoms induced by ZYMV:WK infection did not intensify to severe leaf symptoms and where there were natural outbreaks of severe ZYMV infection, fruits from inoculated plants remained symptomless whilst those from uninoculated plants were severely affected and unmarketable.     

Effect of Globodera rostochiensis on the growth of reciprocal grafts between two potato cultivars of different tolerance in a field trial

The tolerance of plants produced from reciprocal grafts between cv. Cara and cv. Pentland Dell were compared in field plots containing different initial population densities of Globodera rostochiensis. Both stock and scion contributed to the greater tolerance of cv. Cara as compared with cv. Pentland Dell. Ground cover measurements, made by a novel photographic method, showed that a heavy infestation of G. rostochiensis decreased the top growth of all plants: more for those with scions of Pentland Dell than those with scions of Cara. The stock did not initially affect the extent to which top growth was reduced in the heavily infested plots but later measurements showed a progressively increased effect on top growth for plants with stocks of Pentland Dell and a decreased effect for those with Cara. In a destructive harvest 13 wk after planting the relatively small effect of G. rostochiensis on the top growth of plants with Cara scions was not reflected in the tuber yields which were markedly decreased. However, plants with Cara stocks were considerably more tolerant (i.e. tuber yields were affected less) than those with stocks of Pentland Dell. Overall, plants of Pentland Dell scions on Cara root stocks were most tolerant at this stage. Between 13 and 16 wk, when a second destructive harvest was made, plants with Pentland Dell scions were senescent and their yield increased little. In contrast, plants with Cara scions were still growing and their yield increased markedly so that by 16 wk the most tolerant combination was Cara on Cara.     

Factors determining recovery and reversion in mosaic-affected African cassava mosaic virus resistant cassava

The severity and persistence of symptoms of mosaic virus disease were monitored during the first six months of two growing seasons in cassava of the African cassava mosaic virus (ACMV)-resistant cv. TMS 30572 either inoculated by grafting with a mild or severe strain or infected from the planted cutting. Symptomless shoots developed between January and March 1995 in two field trials differing in age by c. 6 months; this recovery occurred during particularly hot weather. Recovery was often only temporary in the plants inoculated with the severe strain and occurred later compared with those inoculated with the mild. In 1996, the weather was cooler and recovery that year was delayed until flowering, c. 7 months after planting, when recovered shoots were often produced from buds in the axils of symptomless leaves produced amongst diseased leaves. Most cuttings taken from the upper parts of diseased plants produced symptomless (reverted) progenies whereas most cuttings taken from the base of diseased plants produced diseased progenies. Reversion seemed to be associated with the recovery that had already occurred in the upper stems of the parent plants.     

Studies on mutagenesis and cross-protection of cauliflower mosaic virus

Three strains of cauliflower mosaic virus (CaMV) designated NVRS, CM4-184 and PK caused respectively severe, moderate and mild reactions in turnip cv. Just Right plants and severe, mild and symptomless reactions in Brussels sprout cv. Fasolt plants. Chlorotic local lesions formed consistently in the leaves of young turnip plants when inoculated with each of the virus strains. Lesions were suitable for infectivity assay of crude and purified preparations of the virus. Three variants of the NVRS strain were isolated by single-lesion transfer after treatment of the virus with nitrous acid (pH 5.0) and two variants were obtained after treating the virus in acetate buffer at the same pH. One of the variants (designated V3) caused symptomless infection in turnip and Brussels sprout plants. In cross-protection tests, Brussels sprout plants infected symptomlessly with the PK, CM4-184 or the V3 strains, subsequently resisted infection by the severe NVRS strain.     

Effects of Inoculation with a Mild Strain of Tomato Aspermy Virus on the Growth and Yield of Tomatoes and the Potential for Cross Protection

Mechanical inoculation of seedlings of 26 tomato cultivars with an attenuated strain of tomato aspermy virus (TAV-M, ATCC # PV361) resulted in development of only mild symptoms, which had largely disappeared by the time the infected plants had begun to set fruit. Yield trials revealed that plants inoculated with TAV-M when transplanted at the 6–8 leaf stage displayed no delay in fruit set and had yields only slightly less than uninoculated plants. Cross protection studies carried out in the greenhouse revealed that TAV-M has the potential to interfere with infection of tomato seedlings by a more severe TAV strain (ATCC # PV127). ‘Pik-Red’ seedlings inoculated with TAV-M showed a significant reduction in symptom expression when challenged with the Blencowe strain of TAV 21 days after the initial inoculation with TAV-M; interference was incomplete at 7 and 14 days.     

Use of resistant and susceptible potato cultivars in the trap cropping of potato cyst nematodes, Globodera pallida and G. rostochiensis

The effects of planting date and growing period of potato cultivars on their efficiency as trap crops for potato cyst nematodes (PCN) were studied. Plots were planted with susceptible or resistant cultivars in April, June and August and these were grown for 5, 6 or 7 wk before removal of the plants by hand lifting. Crops planted in June provided the best overall reductions in PCN population density of up to 95%, with cv. Santé significantly more effective than the other cultivars. Population reductions from the August planting were only slightly less than from planting in June but the tuber yields obtained were much greater: Maris Piper and Maris Bard produced 16.4 and 21.4 t ha^-1 respectively, with 37% and 43% respectively, of a size useful for canning (i.e. between 20 and 40 mm diameter).     

Transmission of Three Viroids Through Seed and Pollen of Tomato Plants

The severe strain of potato spindle tuber viroid (s-PSTV) as well as chrysanthemum stunt (CSV) and cucumber pale fruit (CPFV) viroids were found to be transmitted through seed and pollen of the tomato cvs. Rutgers and Najwcze?niejszy. Plants pollinated with a pollen infected with any of these three viroids became systematically infected. Plant, fruit and seed symptoms of viroid infection were noted on sap- and pollen-inoculated plants and the yield of these plants was reduced. Tomato cv. Rutgers plants grown from infected seeds were symptomless although all three viroids were detected in these plants by bioassay and by electrophoresis on 5% polyacrylamide gel. When DNA complementary to s-PSTV RNA was used for a direct viroid detection in seed samples by spot hybridization technique it hybridized not only with s-PSTV RNA but also with CSV RNA as well as with CPFV RNA.     

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.     

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.     

Cross-Protection and Multiplication of Mild and Severe Strains of TMV in Tomato Plants

The presence of virus having mixed coats has been demonstrated serologically in tomato plants cross-protected with a mild strain (M 11–16) against the severe (UI) strain of tobacco mosaic virus (TMV). The proportion of virus having mixed coats remained constant throughout at ca. 40% even in top leaves where severe symptoms caused by the challenge virus (U1) first appeared. The symptoms caused by the U1 in doubly-inoculated plants occurred in the leaves where:

• (a) the amout of total virus (irrespective of the type of coat) protein was 52% that of U1 strain in singly inoculated plants,
• (b) the multiplication of the severe U1 strain was reduced to only 9% as compared with the leaves singly inoculated with U1,
• (c) the ratio of severe to mild strain was increased to near equality although there was a significant reduction in the multiplication of U1in these leaves,
• (d) the amount of infective U1RNA started to increase.

     

Systemic fungicides in the control of strawberry wilt (Verticillium dahliae)

In a series of experiments between 1970 and 1973 the application of benomyl or thiophanate methyl to field-grown strawberries, planted on Verticillium-mtested land, gave control of wilt for up to 5 months, the duration of control being related to the amount of fungicide applied in the spring. Treatment of inoculated plants grown on in chloropicrin-fumigated soil was effective for at least two seasons. An autumn-planted multi-factorial experiment in heavily-infested soil showed that, to achieve maximum wilt control, it was advantageous to grow cv. Gorella rather than cv. Cambridge Vigour; to drench the runners at planting rather than to dip them in the fungicide suspension; and to use a high concentration (0–075 % a-i-) and large volume (600 ml per plant) for a supplementary treatment in May rather than a lower concentration (0.025 %) ^or smaller volume (400 ml). There were small but significant advantages in applying benomyl rather than thiophanate methyl, and in using 0–2% a.i. suspension at planting rather than 0–05%. No advantage was gained by dividing the spring application into two equal doses applied 2 wk apart. Extrapolation from the logarithmic relation between wilt index and total dose of fungicide applied in the spring suggested that I.I g/plant would have given almost complete control until October; such control had been achieved in an earlier experiment in which 1–2 g/plant was applied. Crop yield in the second year was determined by the treatment applied in the first year, but although these treatments had given significantly better control of wilt in Gorella than in Cambridge Vigour, the greater growth and yield potentials of the latter cultivar had an over-riding effect on crop production; Cambridge Vigour yielded more than Gorella under all chemical treatments, but in the absence of treatment Gorella gave a larger crop than Cambridge Vigour. A proposed regime, entailing spring and autumn applications, is aimed at minimizing the colonization of the plant throughout the year, thus reducing the production of new inoculum and, by limiting the quantity of the pathogen in contact with the systemic chemicals, minimizing the probability of selecting fungicide-resistant variants of V. dahliae.     

Decreased severity of ryegrass mosaic virus after passage through resistant perennial ryegrass

After exposure to infection in the field, the proportion of plants showing distinct symptoms of ryegrass mosaic virus (RMV) was less in perennial than in Italian ryegrass. The perennial ryegrass cv. Mascot had a smaller proportion of plants with symptoms than cv. S.23. Far milder symptoms were induced in test plants by RMV from naturally infected perennial ryegrass plots than from Italian ryegrass plots. Within perennial ryegrass, RMV from cv. Mascot caused milder symptoms than that from cv. S.23. Severe RMV isolated from Italian ryegrass cv. Trident (RMVT) became milder after one passage through cv. Mascot, although not as mild as RMV obtained from field plots of cv. Mascot (RMVM). Families from two highly resistant perennial ryegrass clones and two randomly selected clones of cv. S.23 crossed in all possible combinations varied in symptom severity when inoculated with RMVT but not when inoculated with RMVM. Families inoculated with RMVT also yielded virus which varied in the severity of symptoms induced in test plants, families with severe symptoms yielding severer virus. Thus, much of the variation in the resistance of these clones could be attributed to infection with RMV of differing severity. Resistance was controlled by several genes which were mainly additive in their effect.     

含卫星RNA的黄瓜花叶病毒弱株系的分离鉴定及在病毒病防治上的应用

从豌豆上分离获得黄瓜花叶病毒分离物ＣＭＶＰ１,摩擦接种９科２９种植物,ＣＭＶＰｌ在大多数植物上症状很轻或无任何症状,提纯的病毒颗粒为球形,直径约２８ｎｍ,病毒衣壳蛋白亚基分子量约２７．５ｋＤ,所含核酸有五个组份,即ＣＭＶＰ１含有卫星ＲＮＡ。ＣＭＶＰ１接种三生烟后８－１２天内呈轻花叶,此时组织中病毒含量最高,随后症状消失,去除卫星ＲＮＡ能加重ＣＭＶＰ１在番茄上的症状,因而是卫星ＲＮＡ减轻了ＣＭＶＰｌ的病状。当ＣＭＶＰ１保护接种番茄后攻强毒,番茄发病率低,病情轻,保护率达９０％以上,并有一定的刺激生长作用,还能提早开花４天,植株结果数增多,成熟果实的颜色、形态、品质和重量均正常。ＣＭＶＰ１对烟草亦具有很好的保护效果。保护接种的植株能明显减少强毒株侵染,可减少９０％以上。     

Effects of common scab (Streptomyces scabies) on potatoes

Seed tubers with severe (71% cover) or slight (12% cover) common scab selected from a badly infected crop of cv. Maris Piper were boxed for sprouting in November and February and planted in field experiments at Woburn and Rothamsted in 1980. The severely scabbed seed lost more moisture than the slightly scabbed seed during sprouting and sprouted from a larger number of eyes but total stem numbers were greater on plants from the slightly scabbed seed. Plant growth during the first 6–9 wk after planting was less from the severely scabbed than from the slightly scabbed seed. This was shown by records of ground cover, leaf area index, fresh weights of stems and leaves, total tuber yield (18% lower at 9 wk) and tuber number. Subsequently these differences became non-significant. Seed with slight scab boxed in November produced smaller numbers of progeny tubers than that boxed in February but there were no other consistent effects of boxing date on growth and yield. Scab incidence on progeny tubers was very slight and unrelated to the severity of disease on the seed tubers. It seems unlikely that severe scab infection of seed tubers would significantly decrease tuber yields except perhaps in early potato production.     

Characterization of grapefruit plants (<Emphasis Type="Italic">Citrus paradisi</Emphasis> Macf.) transformed with citrus tristeza closterovirus genes

Grapefruit (Citrus paradisi Macf. cv Duncan) plants were transformed with several sequences from citrus tristeza closterovirus (CTV) that varied in terms of position in the CTV genome and virus strain origin in an attempt to obtain resistant plants. The sequences included the capsid protein gene from three different strains, a nontranslatable version of the capsid protein gene, the replicase (RdRp), the minor capsid protein (p27), a highly transcribed gene of unknown function (p20) and the more conserved 3' end of the genomic RNA. Transgenic plants were generated from all of the constructs, except from the p20 and p27 genes. Southern and Western blot analyses demonstrated that stably transformed grapefruit plants were obtained and that at least some transgenes were expressed. In a first effort at virus challenge, 25 transgenic lines were graft inoculated with a severe strain of CTV. Although some transgenic plants averaged lower titers of virus than controls, there was great variability in titer in both controls and transgenic plants, and all were apparently susceptible to the virus.     

Differences in Endogenous Levels of Gibberellin-Like Substances in Nodules of Phaseolus lunatus L. Plants Inoculated with Two Rhizobium Strains

Lima bean plants (Phaseolus lunatus L.) inoculated with Rhizobium sp. strain 127E14, which lacks constitutive nitrate reductase activity, were significantly taller after 4 weeks of age than plants inoculated with strain 127E15, which contains constitutive nitrate reductase activity. Plants inoculated with either strain responded to application of 5 micrograms gibberellic acid per plant with rapid internode elongation; plants inoculated with strain 127E15 became less responsive to gibberellic acid from 3 to 5 weeks of age, while plants inoculated with strain 127E14 did not. The height of plants inoculated with strain 127E14 was reduced by 20% with application of gibberellin biosynthesis inhibitors to the roots, while height of plants inoculated with strain 127E15 was unaffected.     

Effects of cultural practices, Verticillium nubilum and locality on coiled sprout and yield of early potatoes

Treatments reported to affect the incidence of ‘coiled sprout’ were studied in 1968 with the cultivar Arran Pilot in experiments near Falmouth, Cornwall (F), Rothamsted (R) and near Edinburgh (E). Most coiled stems occurred at E (37%) with similar proportions at R (19%) and F (16%). Seed tubers with the longest sprouts at planting produced most coils at all sites, those with shorter sprouts less and unsprouted seed least. When coiling was assessed on the number of plants, long- and short-sprouted seed tubers were about equally affected at R and E. Sprouts emerged within 45 days at E and 30 days at R. Longer sprouts did not emerge more quickly, but emergence was more than 1 wk later from unsprouted seed at R and only a few days later at E. Harrowing ridges after planting to decrease the depth of soil covering seed tubers hastened emergence by about 1 wk at R and decreased the incidence of coiling; it had little effect at E but tended to increase coiling at F. Seed tubers inoculated with Verticillium nubilum produced more coiled sprouts but this increase was not significant. The highest final yields were achieved at R (43-2 t/ha) where short-sprouted seed outyielded long-sprouted and unsprouted seed, but cultivation treatments and fungus inoculation had no effect. At E (21-2 t/ha) only herbicide treated plots gave a significant effect, outyielding harrowed plots by 8 t/ha. At F (18-2 t/ha) short-sprouted seed outyielded long-sprouted and unsprouted seed, and herbicide treated plots outyielded harrowed plots. When ware yields from individual plants with and without coils from long-sprouted seed were compared at R on 18 June, coiled plants gave about 30% and on 2 July 60% of the yield of non-coiled, but by 16 July 120% yield (20% more); similar trends were shown by short- and by long-sprouted seed at E.     

Incidence, field spread, seed transmission and effects of broad bean stain virus and Echtes Ackerbohnenmosaik-Virus in Vicia faba in eastern Scotland

In eastern Scotland seed-borne infection with broad bean stain virus (BBSV) and/or Echtes Ackerbohnenmosaik-Virus (EAMV) was detected in five of 39 seed lots of field bean in 1975 and in four of 21 commercial crops of field bean or broad bean sampled in 1975 or 1976. Tests failed to detect the main weevil vector of these viruses, Apion vorax, in 1975 and 1976 but Sitona weevils were found in most crops and were numerous in many, reaching maximum numbers in August. No spread of BBSV and EAMV was detected in commercial crops containing seed-borne infection. In experimental field bean crops containing plants manually inoculated with virus, no virus spread was detected in 1975, and only 0–015% uninoculated plants became infected with EAMV in 1976. Sitona, therefore, was an inefficient vector. The percentage of virus infection in seed harvested from field bean plants manually inoculated 3, 5, 7 and 11 wk after emergence in the field was 1–5, 2–7, 0–4 and 0–06 for BBSV and 0–5, 2-1, 0–6 and 0 for EAMV respectively. Seed harvested from unrogued and rogued plots of field bean grown from seed containing 3–4% seed-borne infection produced 0–05% and no infected plants, respectively. Yield losses in field bean plants manually inoculated with virus before flowering were up to 20% but were much greater in plants infected through the seed. Loss in yield was largely caused by a decrease in number of seeds per pod. The absence of A. vorax, the late arrival of Sitona weevils in the crop and their inefficiency as vectors, and the smaller effects of BBSV and EAMV on crop yield than in southern England appear to make eastern Scotland very suitable for the production of bean seed free from BBSV and EAMV.