Aerial dispersal of the potato gangrene pathogen, Phoma exigua var. foveata

A semi-selective medium for isolating the potato gangrene pathogen, Phoma exigua var. foveata, from air is described. Airborne propagules of P. exigua var. foveata were generated for at least 57 min after the start of simulated raindrop impaction on senescent potato stems bearing pycnidia from a natural infection. The number of propagules generated as aerosols was 0·17% of that available for aerosolisation. The aerodynamic particle size of at least 92% of the aerosols was in the range 2·1–4·7 μm in diameter. Using a Casella High Volume Bacterial Sampler airborne propagules of P. exigua var. foveata were detected in the open air during rainfall from September to November at three sites in Southern Scotland. They were recovered at least 800 m down wind from the nearest potato crop. P. exigua var. exigua and P. eupyrena were also recovered from the air during rainfall. Very low numbers of P. exigua var. foveata were occasionally detected in the absence of rain. The role of airborne inoculum of P. exigua var. foveata is discussed in relation to reinfection of virus-tested stocks of seed potatoes derived from stem cuttings.     

Infection of potato stems by the gangrene pathogen, Phoma exigua var. foveata

Limited transport of pycnidiospores of Phoma exigua var. foveata within transpiring stems was demonstrated by isolating from cut mature stems placed for 24 h in a spore suspension. Pieces of shoot dipped in spore suspension and subsequently rooted contained the pathogen which was redistributed as the plants grew but without causing symptoms until after desiccation. The fungus was only rarely detected within stems growing from rotted seed tubers suggesting that spore transport in the vascular system was relatively unimportant in these experiments. Stems were readily infected from spore suspensions applied to unwounded epidermis especially when the inoculum was held in cotton wool and secured with wax film. The progressive colonisation of growing stems was demonstrated by isolations made at different positions on the stem and at different times after inoculation. In field experiments in two years stems were inoculated with four spore concentrations on four dates from late June to late August. After desiccation the extent of pycnidial lesions was greatest from the earliest inoculation date and the largest spore concentration and these differences were reflected subsequently in the incidence of gangrene on damaged progeny tubers. Younger stems may be more susceptible and infection in early summer probably results in extensive growth within the stem before desiccation and thus in massive pycnidial production.     

The significance of tuber damage and inoculum concentration of Phoma exigua var. foveata in the development of gangrene in stored potato tubers

The susceptibility to gangrene infection of wounds of various shapes and depths on potato tubers was studied by inflicting wounds using differently-shaped brass teeth and rods of different diameters. Inoculating wounds with spore suspensions or damaging tubers which had been previously contaminated with Phoma exigua var. foveata or which had been recently lifted from plots of field experiments showed that wounds in which tissue was crushed were most susceptible to infection. Over a wide range of inoculum concentrations and in experiments using several different cultivars the incidence of infection of any wound type was compared to that of the standard severe cut and crush wound. Using a probit transformation a linear relationship was established, the slope of the line indicating the relative susceptibility of the wound. In 1977 and 1978, crops of cv. Pentland Dell were surveyed for damage incidence, inoculum and inoculum potential on arrival at a commercial bulk store. Nets of tubers buried among the tuber bulk were recovered after storage and gangrene incidence compared with damage and inoculum assessments. Inoculum potential and incidence of severe damage both influenced disease development but damage incidence was of greater importance, showing that priority should be given to decreasing damage and to curing to promote rapid wound healing in endeavours to control the disease.     

The spread of Phoma exigua within the potato crop

The incidence of gangrene that developed in tuber samples harvested in a systematic way from potato crops planted with surface sterilized gangrene-free seed surrounding a small plot planted with gangrene-affected seed showed that in two out of the three years marked outward spread of Phoma exigua var.foveata occurred in an uneven manner to about 4 ft (1–2 m) from the central plot. The occurrence of var. foveata was restricted to the immediate area in and around the gangrene infection locus whilst P. exigua var. exigua was more widely and evenly distributed throughout the experimental area. The incidence of infection in and around the central plot varied from year to year and it is suggested that the main method of spread of var. foveata inoculum is by rain splash. Infection by both fungi appeared to be influenced in a similar manner in all three seasons.     

Variation in pathogenicity among field isolates of Phoma exigua var. foveata

Experiments carried out using a point inoculation method to infect potato tubers with Phoma exigua var. foveata demonstrated considerable variation in pathogenicity among field isolates. This variation was unlikely to be due to differential reductions in isolate pathogenicities during axenic culture. However, fresh field isolates generally produced larger lesions than stored isolates. An investigation of the distribution of pathogenicity variation in the fungus revealed that differences among isolates from different lesions taken from the same potato stock were greater than those between stocks, but pathogenicity variation within each lesion isolate was small relative to that between isolates. The importance of using isolates with a high level of pathogenicity, hence recent field isolates, in studies of this pathogen is stressed.     

Seasonal occurrence of the potato gangrene pathogen, Phoma exigua var. foveata, in the open air

Using a Casella High Volume Bacterial Sampler air samples were collected down wind of potato fields in central Scotland when it was raining and when it was dry between December 1978 and October 1982. Airborne propagules of Phoma exigua var. foveata were recovered from the open air during rainfall in most months of the year but the frequency of recovery was greatest in September, October and November. The numbers of colonies of P. exigua var. foveata detected during rainfall were much greater in September and October than during the rest of the year. Low numbers of colonies of P. exigua var. foveata were recovered in the absence of rainfall and their occurrence appeared to be associated with strong winds and dry soil conditions. These latter airborne propagules were detected during all the months except November to February when few samples were made. Sampling simultaneously up wind and down wind of potato fields in September and October demonstrated that the potato crop was the main source of airborne propagules caught down wind but small numbers of P. exigua var. foveata were recovered up wind in four out of the seven experiments.     

Factors affecting the assessment of resistance of potatoes to gangrene (Phoma exigua var. foveata)

Three experiments designed to elucidate the effects and interactions of different inoculation methods, inoculation positions, strains of Phoma exigua var. foveata and clones of potato tubers inoculated were analysed using as data dimensions of the lesions and a more general surface score. The assessment of susceptibility was affected by all these factors. The ranking of varieties for resistance altered considerably depending upon whether the surface area or the depth of lesions was considered. Derivation of more complex scores combining these measurements reduced the relative genetic component and hence would be of no value to potato breeders.     

The Influence of Glycoalkaloids and Impact Damage on Resistance to Fusarium solani var. coeruleum and Phoma exigua var. foveata in Potato Tubers

Potato glycoalkaloids were determined in tubers of a wide range of genotypes in order to study their influence on resistance to storage rots caused by Fusarium solani var. coeruleum and Phoma exigua var. foveata. No relationships were found. This indicates that breeders selection for low glycoalkaloid potato clones is not in conflict with the demands for high levels of resistance to these pathogens. It was also observed that genotypes susceptible to skin-breaking types of, damage were more easily infected than genotypes susceptible to other types of damage.     

Contamination of seed potato tubers grown from stem cutting or microplant-derived tubers by Phoma foveata and Polyscytalum pustulans in Scotland: the influence of site and fungicide treatment at harvest

A survey was made over 5 years of the incidence of contamination by Phoma foveata and Polyscytalum pustulans on 3-year-old virus-tested stem cutting (VTSC) stocks assessed on three occasions during the storage season, and on their daughter tubers at five farms. The contamination occurring on plants grown in small plots from stem-cutting/microplant tubers was also assessed at the farms.
Fungicide treatment soon after harvest reduced the incidence of both pathogens, especially P. pustulans . Tuber contamination was similar for samples collected after fungicide treatment and before planting. The incidence of contamination on daughter tubers, relative to that on the untreated mother tubers, was less with P. foveata but was similar or greater with P. pustulans . Contamination by both pathogens varied between farms and was reflected in the degree of contamination occurring on daughter tubers from stem-cutting/microplant tubers.
The incidence of P. foveata on stems, soil and tubers of seed stocks classified at Foundation Stock grade from various regions of Scotland was assessed over 2 years. The incidence of tuber contamination was correlated ( P < 0.05) with the amount of tuber-borne soil contamination for all comparisons, but was correlated with stem inoculum in only 7 out of 13 comparisons. The largest amounts of stem inoculum of P. foveata were found in stocks in Aberdeenshire.
These results are discussed in relation to the spread of both pathogens to healthy VTSC stocks.     

Persistence of Phoma exigua xar. foveata and Polyscytalum pustulans in dry soils from potato stores in relation to reinfection of stocks derived from stem cuttings

The incidence of Phoma exigua var. foveata and Polyscytalum pustulans in dry soil and dust from potato stores was assessed at 10 farms in Scotland producing virus-tested stocks of seed potatoes derived from stem cuttings (VTSC). Samples were collected on three occasions during 1976: during the storage period (February-March), soon after the potatoes were planted (May) and just before the new crop was lifted and stored (August-October). Both pathogens were detected at all three sampling times but P. exigua var. foveata was more frequently detected at the last one than P. pustulans. Soil and dust from various sites within the stores, such as floors, ledges, graders and boxes, were contaminated by these pathogens. Propagules of both pathogens remained airborne for at least 12–17 min after the floors were swept. P. exigua var. foveata remained viable for at least 16 months in dry soil from tuber surfaces stored at 4–6°C. When VTSC tubers were dusted before planting with a dry store soil contaminated by P. exigua var. foveata the gangrene potential of the progeny tubers was much greater than that from untreated tubers or from tubers treated with an uncontaminated soil. The role of inoculum surviving in dry soil is discussed in relation to reinfection of VTSC stocks.     

The effect of harvest date and duration of wound healing conditions on the susceptibility of damaged potato tubers to infection by Phoma exigua (gangrene)

In experiments extending over 3 yr, King Edward tubers harvested on various dates from early September until early November were uniformly wounded and kept at 15 ^oC and c. 95% r.h. for periods of up to 21 days for wounds to cure. When tubers were subsequently inoculated with Phoma exigua var.foveata and incubated at 5 ^oC for 12 wk, curing for 3 and 7 days decreased gangrene incidence to respectively c. 13 and 4% of the non-cured controls. When tubers were inoculated before curing and immediately after wounding, curing for 7 days decreased gangrene incidence to only c. 68% of the non-cured controls. Curing was also progressively less effective the later the date of harvesting, suggesting that there would be advantages in harvesting potato crops early in the autumn.     

The size distribution of potato tubers and its application to grading schemes

The weight, length, breadth and depth were measured for each of 10 000 potato tubers. These were samples over 3 years of the 10 most commonly grown UK maincrop varieties. Volume, square-mesh riddle size and average projected area were also measured in the final 2 years. A multivariate log-normal distribution was found to give a good fit to the data. Illustrations are presented of how this distribution can be used to assess grading schemes.     

Selected cultural and environmental parameters influence disease severity of dandelion caused by the potential bioherbicidal fungi, Phoma herbarum and Phoma exigua

Selected cultural and environmental variables were investigated for their influence on the efficacy of Phoma herbarum and Phoma exigua to cause disease on dandelion (Taraxacum officinale) under growth room conditions. In both species, mycelial fragments caused significantly greater disease severity on dandelion than spore suspensions. Mycelial age was not an important factor in disease severity caused by P. herbarum, with all cultures causing high disease ratings. However, younger cultures of P. exigua caused the greatest disease severity on dandelion, but significantly less than that caused by P. herbarum. The initial pH of the growth medium (potato dextrose broth) did not affect disease severity caused by either Phoma species. Increasing concentrations of mycelia of P. herbarum were applied to dandelions that were then exposed to various leaf wetness durations. Disease severity increased with increasing leaf wetness duration. For dandelions exposed to no leaf wetness duration, the greater the mycelial concentration, the greater the disease rating. However, for dandelions exposed to all leaf wetness durations, all concentrations of mycelia caused similar disease ratings. As P. herbarum caused high disease ratings on dandelion, it therefore warrants further investigation as a potential bioherbicide for the control of this weed.     

The plastid machinery of apical meristematic cells of potato tubers whose growth is regulated by melaphene

A comparative ultramorphometric study of the plastid machinery of apical meristematic cells of potato (Solanum tuberosum L.) tubers in a normal state (during sprouting) and under treatment with melaphene, a growth regulator of a new generation, at a growth-stimulating concentration was performed. The area of the plastid machinery of cells was found to increase under the effect of melaphene. A stimulatory effect of melaphene on starch accumulation and development of peripheral plastid reticulum in leucoplasts of apical meristematic cells of potato tubers was discovered.     

The influence of alterations in ADP-glucose pyrophosphorylase activities on starch structure and composition in potato tubers

In order to examine whether alterations in the supply of precursor molecules into the starch biosynthetic pathway affected various characteristics of the starch, starch was isolated from potato (Solanum tuberosum L.) tubers containing reduced amounts of the enzyme ADP-glucose pyrophosphorylase (AGPase). It was found that although the type of crystalline polymorph in the starch was not altered, the amylose content was severely reduced. In addition, amylopectin from the transgenic plants accumulated more relatively short chains than that from control plants and the sizes of starch granules were reduced. The starch granules from the transgenic plants contained a greater amount of granule-bound starch synthase enzyme, which led to an increase in the maximum activity of the enzyme per unit starch tested. The K _m for ADP-glucose was, at most, only slightly altered in the transgenic lines. Potato plants containing reduced AGPase activity were also transformed with a bacterial gene coding for AGPase to test whether this enzyme can incorporate phosphate monoesters into amylopectin. A slight increase in phosphate contents in the starch in comparison with the untransformed control was found, but not in comparison with starch from the line with reduced AGPase activity into which the bacterial gene was transformed. Received: 2 February 1999 / Accepted: 25 March 1999     

Decreased amount of reducing sugars in transgenic potato tubers and its influence on yield characteristics

This work focuses on the comparison of field characteristics and amounts of reducing sugars in cold-stored tubers of transgenic plants derived from two potato cultivars. The bacterial gene coding for phosphofructokinase under the tuber-specific promoter was used to support the glycolysis in stored tubers. While the tubers from untransformed control plants steadily accumulated reducing sugars during cold storage, the tubers from transformed plants regardless the genotype were characterized by subsequent decrease in the sugar content. After long period of cold storage the greatest reduction in the reducing sugar content was by more than 60 % compared to control. Before the storage, however, the content of reducing sugars was in 80 % of transgenic lines higher than in control ones. The plants evaluated in field trials for their appearance showed any changes in growth characteristics in about 25 % of the transgenic lines. Despite the introduced modification of sugar metabolism the yield of transgenic plants with normal appearance did not differ significantly from the yield of control plants.     

Methods of measuring the prevalence of Phoma exigua on potatoes and in soil

Methods are described for assessing inoculum of Phoma exigua by directly wounding potato test tubers and by inoculating soil into Arran Banner test tubers or slices. Large samples of tubers were wounded by dropping onto perforated metal sheet or grading on farm riddles, but standardization was achieved by inflicting four uniform wounds with shaped brass teeth. Tubers were then incubated for 12 wk at 5 ^oC. Soil samples to be tested were inoculated into crush wounds on surface-sterilized test tubers or tuber slices which were incubated for 8 wk at 5 ^oC. Assessments were speeded by immersing wounded slices in a mixture of Agrimycin and isopropylphenyl carbamate (IPC) solutions before inoculating, and then incubating for 2–3 wk at 10 ^oC. Although the sensitivity of test tubers is limited by their latent infection, comparisons of the tests in 3–5 years showed that the test tuber methods were frequently more sensitive than direct wounding, especially when amounts of inoculum were small.     

Effects of fungicides applied to seed potato tubers at harvest or after grading on fungal storage diseases and plant development

An assessment was made over 3 years of the efficacy of thiabendazole, 2–aminobutane and a formulation of a mixture of thiabendazole and 2–aminobutane in controlling gangrene, skin spot and dry rot when applied either at harvest or after grading at different dates during the storage season.
After harvest applications, the incidence of gangrene was least on tubers treated with the mixture and generally greatest with thiabendazole. When applied after grading the efficacy of the fungicides was not affected by date of grading. Although 2–aminobutane applications at harvest or after grading were equally effective in controlling gangrene, thiabendazole and the mixture were less effective after grading where gangrene was prevalent in a stock. Thiabendazole and the mixture gave good control of dry rot when applied at harvest but were less effective when applied after grading, particularly late in the storage season. Dry rot was sometimes more prevalent on tubers fumigated with 2–aminobutane, especially those treated after grading. Skin spot was more effectively controlled by the mixture and 2–aminobutane than by thiabendazole but, with applications after grading, the efficacy of all fungicides declined the later the date of grading.
Plants grown from tubers treated with the mixture or its 2–aminobutane glycollate constituent exhibited characteristics associated with genetic variation.