Effect of Co-infection by Fusarium oxysporum and Cowpea Mosaic Virus on the Growth and Colonization of Cowpea Seedlings (Vigna unguiculata [L.] Walp.)

Combined infection of cowpea seedlings (c. v. ‘California Blackeye”) by cowpea mosaic virus (CPMV) and Fusarium oxysporum induced greater losses in leaf area, fresh and dry weights than infection by either pathogen alone. The growth of seedlings infected by F. oxysporum f. sp. tracheiphilum was less than that of comparable seedlings infected by F. oxysporum f. sp. phaseoli. The virus infectivity of extracts of the trifoliate leaves of dual-infected plants was significantly higher than that of comparable extracts from the leaves of plants singly infected with CPMV. The nature of the effects of multiple infection in cowpea is discussed.     

The isolation and identification of rhubarb viruses occurring in Britain

Virus-like symptoms were common in British crops of rhubarb. All plants tested of the three main varieties, ‘Timperley Early’, ‘Prince Albert’ and ‘Victoria’, were virus-infected. Turnip mosaic virus and a severe isolate of arabis mosaic virus (AMV) were obtained from ‘Timperley Early’; and ‘Prince Albert’ contained turnip mosaic virus, cherry leaf roll virus (CLRV), a mild isolate of AMV and, infrequently, cucumber mosaic virus (CMV). The main commercial variety ‘Victoria’ contained turnip mosaic virus, CLRV, a mild isolate of AMV and, infrequently, strawberry latent ringspot virus (SLRV). All the viruses were identified serologically. The rhubarb isolates did not differ markedly from other isolates of these viruses in herbaceous host reactions, properties in vitro or particle size and shape. A rhubarb isolate of CLRV was distinguished serologically from a cherry isolate of the virus. Turnip mosaic virus, CLRV and SLRV, were transmitted with difficulty, but AMV isolates were readily transmitted by mechanical inoculation. Turnip mosaic virus was also transmitted to rhubarb by Myzus persicae and Aphis fabae. CLRV was transmitted in 6–8% of the seed of infected ‘Prince Albert’ and ‘Victoria’ rhubarb and in 72% of the seed of infected Chenopodium amaranticolor. Mild isolates of AMV were also transmitted in 10–24% of the seed of infected ‘Prince Albert’ and ‘Victoria’ plants.     

Characterization of Fusarium oxysporum Isolates from Common Bean and Sugar Beet Using Pathogenicity Assays and Random-amplified Polymorphic DNA Markers

Fusarium wilt is an economically important fungal disease of common bean and sugar beet in the Central High Plains (CHP) region of the USA, with yield losses approaching 30% under appropriate environmental conditions. The objective of this study was to characterize genetic diversity and pathogenicity of isolates of Fusarium oxysporum obtained from common bean and sugar beet plants in the CHP that exhibited Fusarium wilt symptoms. A total of 166 isolates of F. oxysporum isolated from diseased common bean plants were screened for pathogenicity on the universal susceptible common bean cultivar ‘UI 114’. Only four of 166 isolates were pathogenic and were designated F. oxysporum f.sp. phaseoli (Fop). A set of 34 isolates, including pathogenic Fop, F. oxysporum f.sp. betae (Fob) isolates pathogenic on sugar beet, and non‐pathogenic (Fo) isolates, were selected for random‐amplified polymorphic DNA (RAPD) analysis. A total of 12 RAPD primers, which generated 105 polymorphic bands, were used to construct an unweighted paired group method with arithmetic averages dendrogram based on Jaccard's coefficient of similarity. All CHP Fop isolates had identical RAPD banding patterns, suggesting low genetic diversity for Fop in this region. CHP Fob isolates showed a greater degree of diversity, but in general clustered together in a grouping distinct from Fop isolates. As RAPD markers revealed such a high level of genetic diversity across all isolates examined, we conclude that RAPD markers had only limited usefulness in correlating pathogenicity among the isolates and races in this study.     

The effect of light wavelength on the susceptibility of plants to virus infection

Plants exposed for 24–72 h to light of different wavelengths differed in their subsequent susceptibility to virus infection. French bean leaves were less susceptible to infection by tobacco necrosis virus and Nicotiana glutinosa leaves were less susceptible to infection by tobacco mosaic virus when previously exposed to blue or red light than when exposed to green light. These differences were most pronounced at low energy levels. Leaves exposed to each kind of light were less susceptible than those kept in darkness.     

Growth and Nitrogen Fixation Activity of Soybeans in the Presence of Bean Pod Mottle Virus

Pathological and physiological responses associated with the host-microsymbiont interaction of soybeans (Glycine max‘Franklin’, ‘Dyer’, ‘Centennial’, ‘Marshall’, ‘Williams’, ‘Bedford’, and ‘Peking’), which were nodulated with Rhizobium japonicum (USDA strain 3I1b-110) and inoculated with bean pod mottle virus (BPMV) in the greenhouse, varied with the cultivar and the substrate in which the plants were grown. In plants grown in a soil mixture, the virus significantly reduced canopy growth, nodulation, total N, and ureide-N content of most cultivars but symbiotic N₂ fixation was significantly reduced only for ‘Centennial’ and ‘Peking’. In vermiculite, in which the plants depended almost exclusively on symbiotically fixed N_,2, the virus reduced canopy and nodule mass of some cultivars but these reductions were significant only for ‘Franklin’. In plants grown in this substrate, however, the virus had little effect as increases or decreases in measured components of symbiotic N metabolism were nonsignificant except for the increase in leaf ureides in ‘Marshall’. Results suggest that in either soil or vermiculite the symbiotic process in certain soybean cultivars can function at nearly a normal level despite root noduleinfection with this virus.     

Activation of Glycosidases as a Consequence of Infection Stress in Fusarium Wilt of Tomato

Changes of β-1,3-glucanase, chitinase, β-1,4-glucosidase and N-acetylglucosaminidase activity have been investigated in relation to the development of symptoms and colonization by the pathogen in roots, stems and leaves of susceptible (‘Improved, Pearson’) and resistant (‘Improved Pearson VF11’) tomato plants infected by Fusarium oxysporum f. sp. lycopersici. Glycosidase activities increased after inoculation to different extents depending on the plant part and cultivar. Increases were always higher in susceptible than in resistant plants. Changes in the β-1,3-glucanase activity after inoculation were particularly large in stems of infected plants. In contrast, chitinase activity increased more in roots than in stems. The β-1,3-glucosidase and chitinase activity decreased slightly from the basal to the apical third of stems. The trend of changes of the glycosidase activity generally were well related with the severity of disease symptoms and the fungal colonization of basal stem segments. There was no evidence that the increase of glycosidase activity after the infection was directly related with the resistance to Fusarium wilt in tomato.     

Physiology of Rice Tungro Virus Disease: Increased Cytokinin Activity in Tungro-Infected Rice Cultivars

Tungro virus infection decreased the chlorophyll content in the susceptible rice (Oryza sativa) cultivar ‘Taichung Native 1’, while it increased the pigment in the less-susceptible cultivar ‘IR 20’ during the later stages of infection. The virus was recovered from the infected susceptibile cultivar as early as 3 days after inoculation. Relatively low amounts of virus were recovered from the less-susceptible cultivar between 12 and 30 days after inoculation. The senescence of detached leaves of virus-infected less-susceptible cultivar incubated in water in darkness was considerably delayed compared to healthy leaves. The cytokinin activity in the extracts from inoculated less-susceptible cultivar was 69% higher than in the extracts from the healthy plants.     

Pathogenicity of Fusarium solani f. sp. glycines Isolates on Soybean and Green Bean Plants

Green bean plants were grown in a greenhouse in soil removed from a soybean field in 1996 that had a high incidence of soybean sudden death syndrome (SDS). Over a period of 4 weeks, isolations were made from taproot tissue of green bean plants to recover Fusarium isolates. Ten isolates of Fusarium solani were recovered and used to inoculate soybean and green bean plants in the greenhouse. These 10 isolates caused typical SDS symptoms on the soybean plants and caused a root and crown rot on green bean plants. The green bean plants did not develop typical symptoms associated with soybean SDS but, rather, leaves on infected plants showed yellowing and necrosis. Molecular data indicated that these 10 isolates were identical to Fusarium solani f. sp. glycines that cause soybean sudden death syndrome. All isolates were re-isolated from greenhouse-inoculated soybean and green bean plants.     

Biochemical aspects of bean resistance to anthracnose mediated by silicon

This study investigated the effect of silicon (Si) on resistance of bean plants (cv. ‘Peróla’) to anthracnose, caused by Colletotrichum lindemuthianum, grown in a nutrient solution containing 0 (?Si) or 2 mmol Si L^?1 (+Si). The concentration of Si in leaf tissue and the incubation period increased by 55.2% and 14.3%, respectively, in +Si plants in relation to ?Si plants. The area under anthracnose progress curve and the severity estimated by the software QUANT significantly decreased by 32.9% and 27%, respectively, for +Si plants. Si did not affect the concentration of total soluble phenolics. Chitinases activity was higher in the advanced stages of infection by C. lindemuthianum for leaves of ?Si plants. β‐1,3‐Glucanase activity increased after C. lindemuthianum infection, but it was not enhanced by Si. Peroxidase and polyphenoloxidase activities had no apparent effect on the resistance of bean plants to anthracnose, regardless of the presence of Si. The increase in lignin concentration as well as on the phenylalanine ammonia‐lyase and lipoxygenase activities were important for the resistance of +Si plants against anthracnose. The results of this study suggest that Si may increase resistance to anthracnose in bean plants by enhancing certain biochemical mechanisms of defence as opposed to just acting as a physical barrier to penetration by C. lindemuthianum.     

Mixed Infection and Natural Spread of ‘Candidatus Phytoplasma asteris’ and Mungbean Yellow Mosaic India Virus Affecting Soya Bean Crop in India

Suspected phytoplasma and virus‐like symptoms of little leaf, yellow mosaic and witches’ broom were recorded on soya bean and two weed species (Digitaria sanguinalis and Parthenium hysterophorus), at experimental fields of Indian Agricultural Research Institute, New Delhi, India, in August–September 2013. The phytoplasma aetiology was confirmed in symptomatic soya bean and both the weed species by direct and nested PCR assays with phytoplasma‐specific universal primer pairs (P1/P6 and R16F2n/R16R2n). One major leafhopper species viz. Empoasca motti Pruthi feeding on symptomatic soya bean plants was also found phytoplasma positive in nested PCR assays. Sequencing BLASTn search analysis and phylogenetic analysis revealed that 16Sr DNA sequences of phytoplasma isolates of soya bean, weeds and leafhoppers had 99% sequence identity among themselves and were related to strains of ‘Candidatus Phytoplasma asteris’. PCR assays with Mungbean yellow mosaic India virus (MYMIV) coat‐protein‐specific primers yielded an amplicon of approximately 770 bp both from symptomatic soya bean and from whiteflies (Bemisia tabaci) feeding on soya bean, confirmed the presence of MYMIV in soya bean and whitefly. Hence, this study suggested the mixed infection of MYMIV and ‘Ca. P. asteris’ with soya bean yellow leaf and witches’ broom syndrome. The two weed species (D. sanguinalis and P. hysterophorus) were recorded as putative alternative hosts for ‘Ca. P. asteris’ soya bean Indian strain. However, the leafhopper E. motti was recorded as putative vector for the identified soya bean phytoplasma isolate, and the whitefly (B. tabaci) was identified as vector of MYMIV which belonged to Asia‐II‐1 genotype.     

Systemic Infection of Tobacco by Pepper Veinal Mottle Potyvirus (PVMV) Depends on the Presence of Potato Virus Y (PVY)

In single inoculations, both PVY and PVMV replicated in inoculated leaves of Nicotiana tabacum cv. ‘Xanthi nc’ plants, but only PVY infected the tobacco plants systemically, whereas PVMV caused localized infection. A mixed infection by the PVY-To72 and PVMV-type strains was experimentally realized in ‘Xanthi nc’ plants. In the presence of PVY, PVMV migrated systemically into the upper leaves of the tobacco plant, as was proved by back inoculation. It would appear that in tobacco, PVY acts as a “helper” virus, providing PVMV with the necessary component factor for migration. In extracts from the co–infected leaves. Immune Electron Microscopy (IEM) revealed phenotypic mixed particles which contained a mixture of coat proteins of PVY and PVMV. The role of the structural and functional interactions between the two viruses, which enable PVMV to migrate systemically in tobacco plants, is discussed.     

LIGHT QUALITY AND PHOTOREACTIVATION OF PLANTS AND VIRUSES

Visible light of different spectral regions was tested for its ability to reverse three effects of ultra-violet radiation, namely, injury (glazing) of French bean leaves, increased resistance of French bean leaves to infection by the Rothamsted tobacco necrosis virus, and inactivation of potato virus X. The different spectral regions were obtained with colorimetric filters and the filtered and unfiltered light from fluorescent tubes; all three effects were reversed only by regions of wave-lengths shorter than 4700 Å. Thirty minutes of illumination at 300–380 f.c. gave substantial photo-reactivation, but irradiated potato virus X did not become affected by visible light until 30 min. after tobacco leaves were inoculated.     

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.     

Changes in physiology and biochemistry of mottle streak virus infected finger millet plants

A significant reduction in the growth parameters viz., plant height, number of tillers, number of productive tillers, leaves, leaf area, 1000 grain weight and grain yield were observed in the mottle streak virus infected finger millet plants compared to healthy finger millet plants. The germination and vigour of seedlings from the seeds of infected plants were reduced. Physiological changes in finger millet as a result of virus infection were investigated. The chlorophyll pigments ‘a’ and ‘b’ as well as total chlorophyll were reduced due to mottle streak infection. The virus infection led to increased total sugar, starch, soluble protein and phenol contents. The mineral metabolism of infected plants showed a reduction in nitrogen, phosphorus, potassium, magnesium, calcium and iron.     

Molecular Evidence for the Association of Tomato leaf curl Gujarat virus with a Leaf Curl Disease of Phaseolus vulgaris L.

A leaf curl disease with symptoms typical of begomoviruses was observed in bean (Phaseolus vulgaris) at the Main Research Farm of the Indian Institute of Pulses Research, Kanpur, India. Infected plants had severe distortion of leaves and the plants were unproductive. PCR indicated the involvement of French bean leaf curl virus (JQ866297), a recently described Begomovirus, and Tomato leaf curl Gujarat virus (ToLCGV). The full‐length genome of ToLCGV associated with leaf curl disease of bean was 2757 nucleotides long and had maximum identity (97–98%) with seven isolates of ToLCGV (AY234383, AF449999, EU573714, GQ994098, AY190290, FR819708, AF413671) and is designated as Tomato leaf curl Gujarat virus‐(IN:Knp:Bean:2013) (KF440686). To the best of our knowledge, this is the first record of ToLCGV infecting a leguminous host, P. vulgaris.     

Age-related Resistance in Bell Pepper to Cucumber mosaic virus

We demonstrated the occurrence of mature plant resistance in Capsicum annuum‘Early Calwonder’ to Cucumber mosaic virus (CMV) under greenhouse conditions. When Early Calwonder plants were sown at 10 day intervals and transplanted to 10‐cm square pots, three distinct plant sizes were identified that were designated small, medium and large. Trials conducted during each season showed that CMV accumulated in inoculated leaves of all plants of each size category. All small plants (with the exception of the winter trial) developed a systemic infection that included accumulation of CMV in uninoculated leaves and severe systemic symptoms. Medium plants had a range of responses that included no systemic infection to detection of CMV in uninoculated leaves with the systemically infected plants being either symptomless or expressing only mild symptoms. None of the large plants contained detectable amounts of CMV in uninoculated leaves or developed symptoms. When plants were challenged by inoculation of leaves positioned at different locations along the stem or different numbers of leaves were inoculated, large plants continued to accumulate CMV in inoculated leaves but no systemic infection was observed. When systemic infection of large plants did occur, e.g. when CMV‐infected pepper was used as a source of inoculum, virus accumulation in uninoculated leaves was relatively low and plants remained symptomless. A time‐course study of CMV accumulation in inoculated leaves revealed no difference between small and large plants. Analyses to examine movement of CMV into the petiole of inoculated leaves and throughout the stem showed a range in the extent of infection. While all large plants contained CMV in inoculated leaves, some had no detectable amounts of virus beyond the leaf blade, whereas others contained virus throughout the length of the stem but with limited accumulation relative to controls.     

Fungus gnats vector Fusarium oxysporum f.sp. radicislycopersici1

Fungus gnat adults transported Fusarium oxysporum f.sp. radicis-lycopersici from Petri dish culture and infected host plants to the roots and hypocotyls of healthy tomato and bean plants. The source of the fungus did not affect the ability of fungus gnats to transport the fungus to healthy hosts. The presence of fungus gnat larvae in media in which young tomato plants were grown did not increase the incidence of plant infection by the pathogen. Fungus gnat adults appear to aid in the dissemination of F. oxysporum f.sp. radicis-lycopersici.     

Effects of mycorrhizae and chitin-hydrolysing microbes on Vicia faba

The effect of Streptomyces albovinaceus (S-22) and Bacillus sp. (B1) on the growth response, nodulation, nutrition and nitrogenase activities of faba bean (Vicia faba) varieties infected with Glomus mosseae under pot conditions in sterile soil amended with chitin was studied. The growth, nodulation, nutrients content and nitrogenase activity of mycorrhiza-treated plants of Giza-667 were significantly increased compared to untreated ones. Such increases were related to the increase in mycorrhizal root infection. Amendment of soil with chitin alone reduced the growth, nodulation, total nitrogen contents and nitrogenase activities of mycorrhiza-treated faba bean plants (Giza-667) compared to untreated plants. Inoculation of plants with S. albovinaceus or Bacillus sp. significantly increased the level of mycorrhizal roots infection, but addition of chitin to the soil in combination with Bacillus sp. reduced the mycorrhizal infection of faba bean roots. Highest phosphorus contents of faba bean Giza-667 were recorded after G. mosseae inoculation in the presence of all treatments. Similar results were observed for the other varieties. The microbial populations were significantly increased in rhizospheres amended with chitin. Such increases were not in response to the mycorrhizal inoculation. Generally, the microflora of faba bean rhizospheres was increased after treatment with G. mosseae in the absence of chitin amendment alone compared with non-mycorrhizal rhizospheres.     

Effects of biotic and abiotic stress on the growth of three genotypes of Lolium perenne with and without infection by the fungal endophyte Neotyphodium lolii

The combinations of three genotypes of Lolium perenne with and without (i) infection by the fungal endophyte Neotyphodium lolii, (ii) infection by ryegrass mosaic virus and (iii) one of five different forms of abiotic stress were studied in pot experiments in a glasshouse. The five abiotic stress treatments were (i) low pH (compared with ‘optimal’ pH), (ii) cutting plants to a height of 1 cm (compared with 5 cm), (iii) shading (compared with no shading), (iv) cutting plants at 2‐weekly intervals (compared with 6 wk) and (v) low nitrogen applied (compared with ‘high’ nitrogen applied). On average, over the five experiments, the accumulated herbage dry weight was 10% more for N. lolii‐infected plants than uninfected, 22% more for virus‐free plants than infected, and 265% more for ‘unstressed’ plants than for plants with abiotic stress. The effects of N. lolii infection on plant growth when the plants were under abiotic or biotic stress were not consistent.     

Effects of infection with the cabbage black ringspot strain of turnip mosaic virus on turnip as a host to Myzus persicae and Brevicoryne brassicae

When turnip plants with 3–7 leaves were inoculated with cabbage black ringspot virus (CBRSV) on the 3rd rough-leaf, symptoms only appeared on leaves that had been less than 15 mm long at the time of inoculation, although infection decreased the area and both fresh and dry weight of all leaves. Leaves were ‘aged’ by their appearance and placed in Leaf Age Categories (LACs). Leaves with symptoms senesced (‘aged’) prematurely. CBRSV-infection of cv. Green Top White did not change the distribution of populations of Myzus persicae between LACs, but increased the proportion of the plant suitable for colonisation. All suitable LACs were quickly colonised by adult apterae and nymphs. On CBRSV-infected plants the nymphal period was shorter, F₁ adults deposited larvae more frequently and the live body weight and tibial length of the F₂ generation was greater, than on healthy plants. The distribution of Brevicoryne brassicae populations on cv. Green Top White differed from that of M. persicae but was also unchanged by CBRSV-infection. On healthy plants the largest colonies were on mature leaves, so that on virus-infected plants premature senescence shortened the life of the colony. On CBRSV-infected plants the nymphal period was prolonged and the live weight of F₁ and F₂ adult apterae was less than on healthy plants. The differences between the biology of M. persicae and B. brassicae on CBRSV-infected cv. Green Top White were associated with the accelerated senescence of CBRSV-infected leaves. The possibility that CBRSV-infection might reduce the resistance of turnips to aphid infestation was tested. M. persicae and B. brassicae were cultured on two favourable and two less favourable cultivars. No improvement in population growth rate was found when the less favourable host cultivars were infected with CBRSV, but both aphid species weighed less and/or had smaller nymphal populations on cultivars showing the severest symptoms. These results are discussed in relation to the evolution of non-persistent virus transmission by aphids.