Some effects of darkness and partial defoliation on the resistance of sugar beet to downy mildew

Seedlings of two susceptible sugar-beet stocks were grown either in daylight or in darkness for 24 h before or after inoculation with Peronospora farinosa f.sp. betae. Post-inoculation darkness did not significantly affect the percentage of inoculated plants that became infected but increased the percentage of plants on which the fungus sporulated in both stocks. The effects on sporulation of darkness before inoculation were more variable than those of post-inoculation darkness. In the same stocks removal of all leaves, except those in the bud, before inoculation increased the percentage both of plants which became infected and of those on which P. farinosa sporulated. Infection and sporulation were not significantly affected by defoliation in a more resistant stock. Defoliation or darkness may have affected the concentration of certain sugars or amino acids in the bud leaves thereby changing the expression of polygenically-controlled resistance to downy mildew.     

The influence of foliar applications of sugars on the susceptibility of sugar beet to downy mildew

Spraying sugar-beet seedlings in the glasshouse, with 1% or 10% solutions of sucrose, 24 h before inoculation with Peronospora farinosa, significantly reduced their susceptibility to downy mildew. The proportion of inoculated plants that became infected was reduced by spraying with sucrose but the main effect was the inhibition of sporulation. Applications of glucose or fructose also increased the resistance of beet seedlings to P. farinosa. Spraying with sucrose 1 or 2 days before inoculation was much more effective than was spraying shortly before inoculation, or 24 h afterwards, or adding sucrose to the inoculum. Washing sucrose-sprayed seedlings with distilled water 1–2 h before inoculation removed only part of the effect of sucrose on sporulation. Although the mechanism by which applications of sugars affected susceptibility to downy mildew is not understood, the results suggest that the main effects occurred inside the host plant rather than externally. The possible significance of these results, in breeding for resistance to downy mildew and in the control of this disease in the field, is discussed.     

STUDIES ON THE IMPORTANCE OF WILD BEET AS A SOURCE OF PATHOGENS FOR THE SUGAR-BEET CROP

Beet yellows virus, beet mosaic virus, rust ( Uromyces betae (Pers) Lév.), and downy mildew ( Peronospora schachtii Fuckel.) were found to be common in wild beet ( Beta vulgaris s.-sp. maritima L.) growing on the foreshores of south Wales and southern England. The virus diseases were more prevalent in southeast England than in the west, rust more in the west than in the east, and downy mildew is equally prevalent in all regions.
Beet yellows is the most commercially important disease and is more common in sugar-beet crops in East Anglia than elsewhere in Great Britain. There was no evidence that beet yellows spread in East Anglia from wild beet to nearby sugar-beet crops during the springs of 1958 or 1959, and Myzus persicae Sulz., the principal vector of yellows, was rarely found on wild beet growing on the foreshore.
In glasshouse experiments aphids colonized sugar-beet plants watered with tap water in preference to those watered with sea water. Daily watering with sea water made plants unpalatable to aphids within 14 days. Aphids also preferred leaves sprayed with distilled water to those that had been sprayed with sea water. Salt solutions gave results similar to those obtained with sea water.     

Some factors affecting the concentrations of sugars in leaves of inbred sugar-beet lines

In the glasshouse, large differences in concentrations of glucose and sucrose were observed between leaves of inbred sugar-beet lines that are known to differ from each other in resistance to pests and diseases. Differences between these lines in concentrations of fructose, glucose and sucrose were more pronounced in the petiole than in the lamina. The concentrations of glucose and fructose were lower in the first two leaves (primary leaves) than in those produced subsequently (secondary leaves); both types of leaf contained similar concentrations of sucrose. Secondary leaves from plants that had been kept in darkness for 17 h contained less fructose, glucose and sucrose than those of plants kept in sunlight for 5 h. Longer period of darkness lowered the concentrations of monosaccharides in the leaves further but did not affect the sucrose content. Primary and secondary leaves from the same sugar-beet plant often differ in non-race-specific resistance to pests and diseases; and darkness can affect suscetibility of beet to downy mildew and to the aphid Myzus persicae. The results of the sugar determinations therefore support the hypothesis that the concentrations of certain carbohydrates in sugar-beet leaves are important in non-race-specific resistance to pests and diseases.     

Germination of Erysiphe gratninis f.sp. hordei conidia on barley leaves

Germination of Erysiphe graminis f.sp. hordei conidia on leaves of several barley cultivars was studied in the laboratory. On both detached leaves and intact plants, within 48 h of inoculation a higher proportion of conidia had germinated on the basal and middle portions of the adaxial leaf surface than on the corresponding portions of the abaxial surface. Such differences between surfaces were not observed near the leaf tip. Similar results were obtained with all the cultivars and growth stages tested, and with five isolates of E. graminis, and are consistent with the observation that there is usually less powdery mildew on the abaxial than the adaxial surface of barley leaves. With most of the barley genotype/mildew isolate combinations tested, within 48 h of inoculation higher proportions of conidia germinated on seedlings and juvenile plants than on older plants. Inherited characteristics which affect spore germination on the leaf surface may be important factors in the development of adult-plant resistance of barley to powdery mildew, particularly in certain genotypes.     

Intraspecific variation of Myzus persicae on sugar beet (Beta vulgaris)

Differences in inherited resistance among seven sugar-beet stocks had similar effects on Myzus persicae clones representing the range of variation in aphid response to resistant and susceptible sugar beet observed in fifty-eight clones collected between 1969 and 1971. Three sugar-beet stocks were consistently resistant. Statistically significant interactions between beet stocks and aphid clones did not indicate the existence of biotypes with specific abilities to overcome resistance. M. persicae clones differed in their vigour of colonizing sugar beet, irrespective of the differences between beet stocks. The readiness of adult aphids to settle determined the size of aphid population produced and included a component related to the response of the aphid clone to sugar beet as a host, and a component related to the resistance ranking of the beet stock. Breeding sugar beet with resistance to aphids will be simplified, as the results indicate that, at present, differences between aphid biotypes need not be considered a problem.     

Location of Oospores in Buckwheat Seed and Probable Roles of Oospores and Conidia of Peronospora ducometi in the Disease Cycle on Buchwheat

Oospores of Peronospora ducometi, the causal agent of downy mildew of buckwheat (Fagopyrum esculentum), were found in the calyx remnant attached to the seed, on the inside of the seedcoat and in the spermoderm layer between the seedcoat and the endosperm. This constitutes a first report documenting the location of oospores in buckwheat seed. Systemic infection of seedlings occurred from oospore-infested seed. Conidial germination was greater at 14°C than 25°C. Systemic infection also occurred as the result of conidial infection of leaves. It is proposed that primary infection of buckwheat occurs by the germination of seed-borne oospores resulting in systemic invasion of the seedling by the germtubes, and followed by conidial formation on the cotyledons. Secondary infection occurs initially from conidia produced on the cotyledons as a result of the systemic infection from seed and subsequently as the result of repeated infections by conidia produced on leaf lesions as the disease progresses up the plant.     

Conidia of the tomato powdery mildew <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> initiate germ tubes at a predetermined site

The emergence of germ tubes from the conidia of powdery mildew fungi is the first morphological event of the infection process, preceding appressoria formation, peg penetration and primary haustoria formation. Germination patterns of the conidia are specific in powdery mildew fungi and therefore considered useful for identification. In the present study, we examined conidial germination of the tomato powdery mildew Oidium neolycopersici KTP-01 in order to clarify whether germ tube emergence site in KTP-01 conidia is determined by the first contact of the conidia to leaves (as found for the conidia of barley powdery mildew), or alternatively is predetermined and is unrelated to contact stimulus. Highly germinative conidia of KTP-01 were collected from conidial pseudochains on conidiophores in colonies on tomato leaves using two methods involving an electrostatic spore attractor and a blower. In the electrostatic spore attraction method, the conidia were attracted to the electrified insulator probe of the spore collector—this being the first contact stimulus for the conidia. In addition, the blowing method was used as a model of natural infection; pseudochain conidia were transferred to detached leaves by air (1 m/s) from a blower. Thus, landing on the leaves was the first contact for the conidia. Furthermore, conidia were also blown onto an artificial membrane (Parafilm-coated glass slides forming a hydrophobic surface) or solidified agar plates in Petri dishes (hydrophilic surface). Eventually, almost all conidia on the probe and on tomato leaves or artificial hydrophobic and hydrophilic surfaces synchronously germinated within 6 h of incubation, indicating that the first contact of the conidia with any of the aforementioned substrata was an effective germination induction signal. Germ tube emergence sites were exclusively subterminal on the conidia. Moreover, the germ tubes emerged without any relation to the sites touched first on the conidia. Thus, the present study strongly indicates that conidia of O. neolycopersici produce germ tubes at a predetermined site.     

Pathogenicity of the basidiospores of Filobasidiella neoformans

The chronology of Phomopsis ganjae conidia germination and infection of Cannabis sativa leaves was observed with the scanning electron microscope. A-conidia germination approached 100% after 24 h, appresoria initiation began after 36 h; B-conidia germinated by 52 h but were not infective. Four-week-old C. sativa seedlings were more susceptible than 16-week-old plants, males more than females. THC and CBD, extracted and separated via TLC, inhibited P. ganjae conidia germination and hyphal growth.     

Mapping of five resistance genes to sugar-beet powdery mildew using AFLP and anchored SNP markers

Sugar-beet powdery mildew, caused by the fungus Erysiphe betae, now occurs in all sugar-beet growing areas and can reduce sugar yield by up to 30%. Powdery mildew resistant plants from three novel sources were crossed with sugar beet to generate segregating populations. Evaluation of resistance was carried out in artificially inoculated field and controlled environment tests. The resistance level in two of the sources was found to be significantly higher than that in currently available sugar-beet cultivars. AFLP analysis was used in combination with bulked segregant analysis to develop markers linked to the resistant phenotype in each population. Five dominant major resistance genes were identified and assigned the proposed symbols Pm2 to Pm6. Pm3 conferred complete resistance to powdery mildew; the other genes conferred high levels of partial resistance. From the use of anchoring SNP markers, two genes were located to chromosome II and three to chromosome IV. Two of the genes on chromosome IV mapped to the same location and one of the genes on chromosome II mapped to the same region as the previously identified Pm1 gene. With the availability of these genes there is now excellent potential for achieving durable resistance to sugar-beet powdery mildew, thus reducing or obviating the need for chemical control.     

Effects of Heat Shock on Photosynthetic Properties,Antioxidant Enzyme Activity,and Downy Mildew of Cucumber (Cucumis sativus L.)

Heat shock is considered an abiotic stress for plant growth, but the effects of heat shock on physiological responses of cucumber plant leaves with and without downy mildew disease are still not clear. In this study, cucumber seedlings were exposed to heat shock in greenhouses, and the responses of photosynthetic properties, carbohydrate metabolism, antioxidant enzyme activity, osmolytes, and disease severity index of leaves with or without the downy mildew disease were measured. Results showed that heat shock significantly decreased the net photosynthetic rate, actual photochemical efficiency, photochemical quenching coefficient, and starch content. Heat shock caused an increase in the stomatal conductance, transpiration rate, antioxidant enzyme activities, total soluble sugar content, sucrose content, soluble protein content and proline content for both healthy leaves and downy mildew infected leaves. These results demonstrate that heat shock activated the transpiration pathway to protect the photosystem from damage due to excess energy in cucumber leaves. Potential resistance mechanisms of plants exposed to heat stress may involve higher osmotic regulation capacity related to an increase of total accumulations of soluble sugar, proline and soluble protein, as well as higher antioxidant enzymes activity in stressed leaves. Heat shock reduced downy mildew disease severity index by more than 50%, and clearly alleviated downy mildew development in the greenhouses. These findings indicate that cucumber may have a complex physiological change to resist short-term heat shock, and suppress the development of the downy mildew disease.     

Powdery mildew of tobacco (Erysiphe cichoracearum DC.)

Percentage germination, and growth of hyphae from single conidia of Erysiphe cichoracearum DC., were measured on leaf discs from topped and intact tobacco plants, grown in aerated nutrient solutions consisting of basal medium plus large or small amounts of potassium. The effect of supplying sodium was also studied. Discs were incubated on water and on 10% sucrose solution. Changes in free amino nitrogen and carbohydrate in comparable uninfected leaf discs, before and after incubation, were also measured. Potassium deficiency resulted in more free amino nitrogen and soluble carbohydrate and less insoluble carbohydrate, per cm.² of leaf. Spore germination was not greatly affected by treatments, though it was usually less on discs from potassium-deficient leaves. The pathogen grew slower on potassium-deficient leaf discs, whether they were incubated on water or on sucrose. Incubating discs from some leaves on sucrose, compared with water, gave greatly increased sugar content and less fungal growth; discs from other leaves had a much smaller increase in sugar, and hyphal length was similar to that on discs incubated on water. Sodium, when potassium was scarce, increased potassium deficiency symptoms, free amino nitrogen and sugar content, and resistance to powdery mildew.     

Re-consideration of Peronospora farinosa infecting Spinacia oleracea as distinct species, Peronospora effusa

Downy mildew is probably the most widespread and potentially destructive global disease of spinach (Spinacia oleracea). The causal agent of downy mildew disease on various plants of Chenopodiaceae, including spinach, is regarded as a single species, Peronospora farinosa. In the present study, the ITS rDNA sequence and morphological data demonstrated that P. farinosa from S. oleracea is distinct from downy mildew of other chenopodiaceous hosts. Fifty-eight spinach specimens were collected or loaned from 17 countries of Asia, Europe, Oceania, North and South America, which all formed a distinct monophyletic group. No intercontinental genetic variation of the ITS rDNA within Peronospora accessions causing spinach downy mildew disease was found. Phylogenetic trees supported recognition of Peronospora from spinach as a separate species. Microscopic examination also revealed morphological differences between Peronospora specimens from Spinacia and P. farinosa s. lat. specimens from Atriplex, Bassia, Beta, and Chenopodium. Consequently, the name Peronospora effusa should be reinstated for the downy mildew fungus found on spinach. Here, a specimen of the original collections of Peronospora effusa is designated as lectotype.     

SUGAR-BEET YELLOWS: FURTHER STUDIES ON VIRUSES AND VIRUS STRAINS AND THEIR DISTRIBUTION IN EAST ANGLIA, 1958-59

Experiments have shown that, as in the years 1955-57, two yellowing viruses, beet yellows virus (SBYV) and sugar-beet mild yellowing virus (SBMYV), were present in commercial sugar-beet crops in East Anglia in 1958 and 1959. The evidence that they are not closely related viruses has been confirmed. In both years the prevalence of the two viruses was estimated by aphid transmissions from yellowed sugar-beet leaves to healthy sugar beet and Chenopodium capitatum seedlings in the glasshouse, and in 1959 additionally by examination of symptoms on field plants. SBMYV was more common than SBYV over the whole region in 1958, but in 1959 SBYV was slightly more prevalent than SBMYV. In both years SBYV was found more often in the southern than in the northern parts of the region. The results described in this paper suggest that breeding for tolerance to SBMYV may be at least as important economically in East Anglia as breeding for tolerance to SBYV. A wide range of SBYV strains was present in East Anglia in 1959, most of the strains being those which caused severe symptoms in sugar beet and C. capitatum.     

Antifungal Activity of Aqueous Extracts of the Leaves of Cowparsnip and Comfrey

We found that extracts from the leaves of medicinal comfrey and cowparsnip strongly inhibit the germination of Erysiphe graminisconidia and uredospores of Puccinia graminis. Spraying wheat seedlings with these extracts, in contrast to the irrigation of soil, markedly diminished infection in plants with powdery mildew. Antifungal activity in vitroand protective activity (when plants were sprayed) correlated with the level of phenolic compounds in these extracts. Experiments with healthy plants have demonstrated that the photosynthetic apparatus of wheat plants is stimulated by extracts. Spraying seedlings with the extracts resulted in an increased rate of O₂evolution calculated per unit of chlorophyll, an increase in the ratio (F_M– F_T)/F_Tin the experiments that recorded slow fluorescence induction, an increase in the relative light intensity of band A, and a decrease of relative intensity of band Cin experiments with thermoluminescence of wheat leaves. These results provide evidence that the protective activity of comfrey and cowparsnip extracts is associated with their action on the pathogenic fungus and with the activation of natural defense reactions of the host plant.     

Comparison of Different Inoculation Techniques to Screen Resistance of Pea Lines to Downy Mildew

Pea lines with partial resistance to downy mildew were tested with different inoculation techniques. Lines with superior resistance to leaf and pod infection in the field also show superior resistance to artificial inoculation of germinating seeds or seedlings at different ages in greenhouse experiments. Increased resistance of older seedlings was found for all genotypes including the susceptible control. Juvenile leaves at the top of the seedlings plants at time of inoculation, developed the strongest symptoms. Selection for partial resistance can probably be carried out in the greenhouse to improve resistance of adult plants in field.     

Field experiments on sugar-beet downy mildew (Peronospora farinosa)

The numbers of sugar-beet plants infected with downy mildew in experimental crops increased slowly over a period of 8–15 weeks from late May. Few new infections appeared after the middle of August, although less than half the plants were then infected. Most infected plants recovered, and recovery was most rapid and complete in young plants, irrespective of the weather. Plants infected when young produced small roots, but their sugar content and juice purity at harvest was similar to those of uninfected plants. Plants first infected in late July and early August had the smallest sugar content and most juice impurities.     

Effects of soil water level on the development of adult plant resistance to powdery mildew in barley

Barley grown in dry soil developed greater adult plant resistance (APR) to powdery mildew (Erysiphe graminis DC. f. sp. hordei Mérat) than barley grown in wet soil. Conidial germination and appressorium formation were less, and fungal development between formation of appressoria and elongating secondary hyphae on upper leaves was inhibited, when adult plants were grown in dry soil. Mildew colonies expanded more slowly on leaves of adult plants than on leaves of seedlings, especially if adult plants had grown in dry soil. APR was reduced if plants, previously grown in dry soil, were well watered more than 32 h before inoculation. Conidia originating from plants grown in dry soil had a lower solute potential and greater ability to infect plants grown in dry but not wet soil than conidia originating from plants grown in wet soil. APR could not be attributed simply to increased cell wall or cuticle thickness, nor to lowered leaf solute potentials, as has sometimes been suggested for powdery mildew diseases. Increasing plant age and water stress induced increases in cell wall and cuticle thickness, but these changes did not always coincide with changes in disease resistance. Increasing plant age and water stress also lowered leaf solute potentials but fungal solute potentials were lower than leaf solute potentials and, more importantly, were lower than leaf water potentials. Thus, fungal growth was not limited by the availability of water from the host during penetration and hyphal establishment. It is suggested that resistance levels may be determined not by the thickness of epidermal structures, nor by lowering of solute potential per se, but by specific substances harmful to the fungus which accumulate in either cell wall, cuticle or sap, and whose concentration is dependent on the age and water stress of leaves.     

Plant Growth‐promoting Fungus Penicillium oxalicum Enhances Plant Growth and Induces Resistance in Pearl Millet Against Downy Mildew Disease

Plant Growth‐promoting Fungus (PGPF) Penicillium oxalicum was isolated from rhizosphere soil of pearl millet and was tested for its ability to promote growth and induce systemic resistance in pearl millet against downy mildew disease. The fungal isolate P. oxalicum UOM PGPF 16 was identified as P. oxalicum using ITS sequencing and morphological analysis and sequence was deposited at NCBI with accession number KF150220. Pearl millet susceptible seeds were treated with three different inducers (CS, CF and LCF) of PGPF P. oxalicum and all the inducers significantly reduced the downy mildew disease and enhanced plant growth. Among the inducers tested, CS treatment recorded highest seed germination of 91% and 1427 seedling vigour followed by LCF and CF treatments. The vegetative growth parameter and NPK uptake studies under greenhouse conditions revealed that the CS treatment of P. oxalicum remarkably enhanced the parameters tested when compared to control plants. A significant disease protection of 62% and 58% against downy mildew disease was observed in plants pretreated with CS of P. oxalicum under greenhouse and field conditions, respectively. The spatio‐temporal studies revealed that inducers P. oxalicum required a minimum of 3 days for developing maximum disease resistance which was maintained thereafter. The maximum Peroxidase (POX) activity (62.7 U) was observed at 24 h in seedlings treated with CS of PGPF P. oxalicum and the activity gradually reduced at later time points after pathogen inoculation. Chitinase (CHT) activity was significantly higher in inducer treated seedlings when compared to control seedlings inoculated with pathogen after 48 h and remained constant at all time points.     

The influence of temperature on the development and yield of maize in Britain

Sugar beet seedlings were evaluated for resistance to downy mildew by exposing them to natural infection in a glasshouse maintained at a high relative humidity (c. 95–98%). Exposure to natural infection for at least 3 days was required for adequate infection levels which occurred in the majority of tests; failure in others was related to decreases in relative humidity during clear weather. Seedling disease symptoms were different from those of mature plants. In most instances there was adequate differentiation among seedlings in populations with low, but worthwhile, levels of variation for resistance. The application of the method in selection programmes for resistance is discussed.