Verticillium dahliae (Kleb.) Infecting Pumpkin Seed

This study investigated the natural occurrence of Verticillium dahliae (Kleb.) infection in pumpkin (Cucurbita pepo L.) seed. The mean incidence of infection was found to be 21.0%. Isolates recovered from seeds were pathogenic to pumpkin (cultivar ‘Jamaican squash’). Surface sterilization by immersion in 0.6% sodium hypochlorite for 20 min eradicated V. dahliae from infected pumpkin seeds without affecting germinability. Plating of seed components revealed that the fungus was present in the seed coat but not in the embryo or cotyledons. In a growing‐on test, 25% of 6‐week‐old plants grown from untreated seeds were infected. Germination and production of normal seedlings were unaffected by V. dahliae infection of seeds. Verticillium dahliae in pumpkin seed was found to be external and transmissible to plants. The findings of this study are important in devising disease control strategies.     

Transfer of Alternaria macrospora from Cotton Seed to Seedling: Light and Scanning Electron Microscopy of Colonization

Alternaria macrospora is transferred from an infected seed, through the plant stem to the cotyledon and sporulates on its surface. However, at the beginning of the process, there is no difference between resistant and susceptible plants. In later stages, lower levels of fungal mycelium and sporulation were detected on the seedlings of a resistant cultivar. The fungus grew inside the plant from the underground to the upper parts. The mycelia which originated from surface spores could penetrate, through stomata and result in leaf colonization. A. macrospora sporulated ultimately on plant surfaces whereas the internal leaf tissue was colonized by mycelium only. This study suggests that cotyledon infection is a result of seed infection.     

Transmission of Alternaria macrospora in Cotton Seeds

Alternaria macrospora was isolated from seeds only after the natural opening of the bolls and exposure of seeds to an environment in which the fungus was present. The fungus lacks the ability to penetrate the boll wall and reach the seed site. Attempts to isolate the pathogen from seeds of immature bolls at different developmental stages failed. Internal infection by slow injection resulted in seed infection and partial shedding of the injected plant parts which was high in buds and decreased with the ripening to mature bolls. Severity of seed infection was not dependent on either inoculum level, boll physiological age or even if the boll itself was not diseased. Infection of flowers under field conditions caused flower shedding. Naturally infected seeds or inoculated seeds with inoculum levels of 100 spores/ml and above resulted in diseased cotyledons, the incidence of which was, for inoculated seeds, positively correlated with inoculum level. A small difference was observed between cultivars in susceptibility to artificial inoculation at the cotyledon stage. A. macrospora survived on commercial cotton seeds and on post-season plants left growing at the field edges. Survival in plant debris under field conditions was minimal and may only have a minor effect on field reinfestation.     

Potato coiled sprout in relation to seed tuber storage treatment and to infection by Verticillium nubilum Pethybr

Inoculation of seed potato tubes with Verticillium nubilum produced infection on the underground stem bases to subsequent growth in the form of brown lesions, often accompanied by longitudinal or transverse cracking. Some inoculation tests provided further evidence of a causal relationship between infection with V. nubilum and coiled sprout, but it was not possible to define the conditions in which the fungus gave rise to coiling. Seed tubers sprouted in light before planting produced more coiling than unsprouted or desprouted tubers. Sprouting had this effect whether V. numbilum was present or not and coiling induced by sprouting and that induced by V. nubilum occurred independently. The increased coiling of stem bases from sprouted tubers was often accompanied by fasciation, whereas this symptom was not usually associated with the coiling from unsprouted or desprouted seed tubers inoculated with the fungus. Brown lesions associated with V. nubilum were readily distinguishable from those of Rhizoctonia solani but not from those caused by Oospora pustulans. Neither of these fungi was found to cause coiling. Different varieties of potatoes showed differences in the incidence of coiling in response to sprouting treatment. There was no apparent differences in varietal response to infection by V. nubilum.     

The effect of elevated temperatures and their duration on the incidence of hollow heart in pea seeds

Plants of Pisum sativum cv. Pania were exposed to high temperature (35 ^oC day/25 ^oC night) at three stages in development. These were: (i) at process crop maturity, when the seed moisture content (m.c.) was 80%, i.e. about 10 days prior to pod wrinkle, (ii) at a seed m.c. of 70 to 80%, about the onset of pod wrinkle, and (iii) when the seed m.c. was 50 to 60%, 10 days after the onset of pod wrinkle. At each stage of development the high temperature treatment was imposed for 2,4,6, 8 or 10 days. Plants exposed to high temperature at seed m.c. of 70 to 80% produced seed with the highest incidence of hollow heart and at all stages of development the incidence increased with the length of exposure to the high temperature. Where plants at the development stage of pod wrinkle (seed m.c. 70–80%) were exposed to a range of temperature regimes for 5 days, over 20% of the seeds had hollow heart when the mean day/night temperature was 25 ^oC. Above a mean temperature of 25 ^oC, the percentage of affected seed increased with increasing day or night temperature. Over 80% of the seeds had hollow heart symptoms after 5 days exposure to a daily mean temperature of 32-5 ^oC. The severity of symptom development was not related to the proportion of seeds with the disorder and, under laboratory conditions, seed germination was not affected.     

Studies on the etiology of black rot, Stemphylium radicinum (Meier, Drechsl. & Eddy) Neerg., and leaf blight, Alternaria dauci (Kühn) Groves & Skolko, on carrot crops; and on fungicide control of their seed-borne infection phases

Of nineteen commercial samples of carrot seed in use at Wellesbourne in 1963, eight were found to be infected with both Stemphylium radicinum and Alternaria dauci and five with S. radicinum alone. Both fungi caused damping-off of seedlings and when carrots were grown at high densities A. dauci caused severe foliar infection. Neither fungus gave rise to marked infection of ware crops at normal spacings but, when seed infected with S. radicinum was sown in the autumn, the resulting seed crop was heavily infected by the succeeding autumn. Spring-sown carrots grown in soil that had borne infected crops or contained debris infected with either fungus showed little or no infection in the autumn or after storage for 3 1/2 months; autumn-sown or spring-planted (steckling) carrots put into ground infected with S. radicinum showed severe infection in the following autumn. This suggested that ware-crop carrots were resistant to infection from the soil whereas the seed crop was not. Both fungi were eradicated from seed by a 24 hr. soak at 30d? C. in a 0.2% thiram suspension, but were not completely eliminated by seed dust treatments with a range of fungicides.     

The response of tomato seedling roots to infection by Verticillium albo-atrum or Fusarium oxysporum f. sp. lycopersici

The response of seedling roots of near-isogenic tomato varieties to infection by Verticillium albo-atrum or Fusarium oxysporum f. sp. lycopersici was investigated. Studies of the infection of seedling roots not artificially damaged indicated that there was an extra-vascular expression of resistance towards V. albo-atrum but not to F. oxysporum. Roots of resistant tomato seedlings infected by V. albo-atrum contained the fungus in the epidermis and outer cortex while susceptible roots became heavily colonised. Observations made by transmission electron microscopy showed that the fungus appeared to be abnormal in growth and appearance in the epidermal and cortical cells of resistant seedling roots but normal in susceptible roots. Two preformed antifungal terpenoids were detected in seedling roots in greater amounts in resistant that in susceptible varieties. The possible mechanisms of seedling root resistance to vascular wilts are discussed.     

Relationship of phenotypic and genetic variation in Plantago lanceolata to disease caused by Fusarium moniliforme var. subglutinans

Summary Naturally established individuals of Plantago lanceolata with the inflorescence disease caused by Fusarium moniliforme var. subglutinans had more inflorescences and were more likely to be male-sterile than healthy plants. Half-sib families planted in the field varied in the percentage of diseased plants, the number of inflorescences per plant, the incidence of male-sterility, and the pattern of inflorescence phenology. The rankings of families with respect to disease incidence was, however, not simply related to their reproductive phenotypes. Plants derived from field genotypes with a history of disease were slightly more likely to become diseased than plants derived from healthy genotypes. Inflorescence infection was more severe on plants derived from genotypes with a known history of disease. Since the fungus reduces seed production in the plants it infects, differential incidence of disease based on plant phenotype and genotype may have ecological and evolutionary consequences for the host population.     

The influence of the testa, damage and seed dressing on the emergence of groundnut (Arachis hypogea)

The effects of damaging the testa and the application of seed dressings were examined in field trials on several short-and long-season cultivars of groundnut (Arachis hypogea) with differently-pigmented testas at Samaru, Nigeria, in 1967. There was a high correlation between the number of seedlings which emerged in these trials and the resistance or susceptibility of the seed to invasion by the fungus Aspergillus flavus, as assessed by laboratory tests. When the testa was undamaged the emergence of white (susceptible) seed was only 50% while that of coloured (resistant) seed was between 95 and 98%. Damage to the testa by scratching greatly decreased emergence. The application of seed dressing increased the emergence of susceptible seed and also restored the emergence of scratched seeds to the level of undamaged seeds. The effect of the complete removal of the testa was not counteracted by seed dressing; naked seeds, with and without dressing, gave c. 10% emergence. The importance of pigment in the testa, the condition of the seed and the effects of seed dressing are discussed.     

Seed cleaning behavior by tropical ants and its anti-fungal effect

In tropical forests, ants frequently consume fruit pulp around seeds of vertebrate-dispersed plants, which protects the seeds from infection by fungus and pathogens. Seed cleaning behavior by omnivorous ants was observed in the secondary forests of Bogor botanical garden in West Java. Fruit pulp around the seeds of the rambutan Nephelium lappaceum was completely removed by Pheidole plagiaria, Anoplolepis gracilipes, and other ants. When seed cleaners were excluded, however, many seeds were attacked by fungus and died. Seeds that were cleaned by hand, but not by ants, were similarly attacked. Field experiments on seed cleaning by P. plagiaria revealed that the anti-fungal effect was not merely caused by removal of fruit pulp, and that seed cleaning reduced the development of spores and hypha of fungi. We suggest that the workers apply anti-fungal substances to the fruit pulp and seed surface.     

Effects of Seedbed Management on Blast Development in Susceptible and Partially Resistant Rice Cultivars

Four rice cultivars, susceptible or partially resistant to Pyricularia grisea were evaluated for their apparent infection rates (r) and for terminal severity values in seedlings grown in seedbeds. Susceptibility of partially resistant cultivars decreased when seeded in wet seedbed compared to those grown in raised or upland seedbed. The degree of blast reduction due to flooding varied with rice genotype, seeding rate, and environmental conditions. Flooding was effective in managing seedling blast of partially resistant cultivars but not of susceptible cultivar. Use of a lower seed rate in comparison to a higher seed rate also led to reductions in r-values and terminal disease severities. However, in case of the susceptible cultivar IR50, effect of seed rate was more pronounced, when low amount and short duration of rainfall occurred. Flooding of wet beds with low seed rate further reduced the blast incidence. Results of this study suggest that with appropriate use of water, seed rate, and partial host resistance there are prospects for effective control of leaf blast in the tropics.     

The occurrence of Aspergillus flavus in vegetative tissue of cotton plants and its relation to seed infection

Twenty-seven mature cotton bolls with Aspergillus flavus Link colonies naturally occurring on the surface of the boll or lint were collected in the field in Arizona along with their subtending stems and peduncles. Bolls inoculated through the carpel wall 30 days after anthesis were allowed to mature in the field and were collected in the same manner. The seed and stem and peduncle sections of each boll were surface-sterilized, plated on agar media and observed for A. flavus. Seventy-eight percent of the naturally contaminated bolls with A. flavus in the seed also had the fungus in the stem and peduncle, whereas only 31% of the naturally contaminated bolls with no A. flavus in the seed had the fungus in the stem or peduncle. This difference was significant (P=0.0125), indicating a positive relationship between seed infection and stem and peduncle infection. All of the bolls inoculated through the carpel wall had A. flavus in the seed, but only 11% of the stem and peduncle sections were infected, indicating that the fungus does not readily grow downward from the boll into the supporting stem or peduncle.This unidirectional pattern of movement (upward) was further substantiated in greenhouse experiments where cotton seedlings were inoculated at the cotyledonary leaf scar with A. flavus and plants were sequentially harvested, surface sterilized and plated. Aspergillus flavus was isolated from the cotyledonary leaf scar, flower buds, developing bolls, and stem sections in the upper portion of the plant. It was never isolated from roots or stem sections below the cotyledonary node, again indicating that the fungus does not readily move downward through the plant.     

Intermittent wetting of soils at high temperature reduces survival of the take-all fungus

Two pot experiments using naturally infested soil and a range of watering regimes were conducted to study the possible effect of level and frequency of wetting of hot soil (to simulate the period between growing seasons in Western Australia) on inoculum of the take-all fungus (Gaeumannomyces graminis var.tritici). In combination with the high soil temperatures, all watering regimes reduced infectivity and propagule number of the take-all fungus, this reduction being absent in dry soils.     

Studies on the seed-borne phases of dark leaf spot Alternaria brassicicola and grey leaf spot Alternaria brassicaeof brassicas

Tests in Britain on samples of basic and commercial Brassica oleracea seed between 1976 and 1978 showed that many lots were infected with Alternaria brassicicola. A. brassicae was uncommon in basic seed in these years and in commercial seed harvested in 1976 and 1977 but was frequent in seed harvested in 1978. Most affected seeds were contaminated by surface-borne spores and mycelium of A. brassicicola but many were internally infected by the fungus situated within the seed-coat and in some seeds in the embryo tissues. Superficial contamination by the fungus declined rapidly after 2 yr in cabbage seeds stored at 10 °C, 50% r.h. but internal infection persisted for up to 12 yr. In some samples, internal infection was commonly associated with small shrivelled seeds. Surface contaminated and internally infected seeds transmitted the disease but seedling infection was more closely correlated with the latter.     

Some observations on assessing Phoma betae infection of sugar-beet seed

Near-ultraviolet or ‘black’ light applied continuously from the start of incubation, facilitates tests for Phoma betae on sugar-beet seed by stimulating the production of pycnidia and restricting mycelial growth of P. betae and other fungi. Pretreatment of the seed with dilute sodium hypochlorite decreases the number of seeds with P. betae by removing superficial infection, but some of this is of significance in the field. Rubbing beet seed also decreased counts of P. betae in the laboratory and increased field emergence, primarily by removing the fungus. Griseofulvin sprayed on beet-seed plants either 2 wk or 2 days before harvest significantly decreased seed infection with P. betae, but not to a level at which further seed treatment could be omitted.     

Control of silver scurf (Helminthosporium solani) disease of potato with benomyl and thiabendazole

Benomyl and thiabendazole, applied as dusts to seed potatoes before chitting (sprouting), reduced the incidence of silver scurf disease on the progeny at lifting and during subsequent storage. Treatment of seed tubers 4 months prior to planting almost completely suppressed sporulation during storage, even under conditions very favourable for the growth of the fungus. The fungicides appeared not to act systemically because developing tubers grown from treated seed were just as susceptible to infection after inoculation as those from untreated. Control was due to effective inhibition of sporulation on the surface of diseased seed tubers after planting. There was no reduction in the amount of disease in subsequent generations of potato crops without further fungicidal treatment. Seed virtually free from silver scurf, produced from seed treated with fungicide in the previous year, yielded progeny with as much disease as progeny from untreated seed stocks.     

Transmission of Pepino mosaic virus by the Fungal Vector Olpidium virulentus

Transmission of Pepino mosaic virus (PepMV) by the fungal vector Olpidium virulentus was studied in two experiments. Two characterized cultures of the fungus were used as stock cultures for the assay: culture A was from lettuce roots collected in Castellón (Spain), and culture B was from tomato roots collected in Murcia (Spain). These fungal cultures were maintained in their original host and irrigated with sterile water. The drainage water collected from irrigating these stock cultures was used for watering PepMV‐infected and non‐infected tomato plants to constitute the acquisition–source plants of the assay, which were divided into six different plots: plants containing fungal culture A (non‐infected and PepMV‐infected); plants containing fungal culture B (non‐infected and PepMV‐infected); PepMV‐infected plants without the fungus; and plants non‐infected either with PepMV and the fungus. Thirty‐six healthy plants grouped into six plots, which constituted the virus acquisition–transmission plants of the assay, were irrigated with different drainage waters obtained by watering the different plots of the acquisition–source plants. PepMV was only transmitted to plants irrigated with the drainage water collected from PepMV‐infected plants whose roots contained the fungal culture B from tomato with a transmission rate of 8%. No infection was detected in plants irrigated with the drainage water collected from plots with only a fungus or virus infection. Both the virus and fungus were detected in water samples collected from the drainage water of the acquisition–source plants of the assay. These transmission assays demonstrated the possibility of PepMV transmission by O. virulentus collected from tomato crops.     

Bioassay for in vitro differentiation of pineapple cultivar resistance levels to heart rot disease

Summary An in vitro bioassay to differentiate pineapple plant resistance levels to Phytophthora nicotianae var. parasitica (heart rot disease) is deseribed here. Conditions to cause death of in vitro-cultured plants were defined using a cultivar previously found to be susceptible to this fungus in our Field-Grown Pineapple Germplasm Bank (ev. Smooth Cayenne Serrana). The effects of zoospore concentration, inoculation technique, and disease progress during the course of time after infection were evaluated. The highest rates of plant death were observed with the use of 10⁸ zoospores ml⁻¹, and the inoculation technique of needle-mediated leaf base wound. One hundred percent plant death was observed at 144h after infection. Different susceptible varieties along with a resistant pineapple relative were additionally compared. In vitro results confirmed previous observations obtained under field conditions. The protocol described here may be used for early selection (in vitro) of new pineapple genotypes showing resistance to this fungus. At present, this protocol is extensively used in the Biotechnology-assisted Cuban Program for Pineapple Breeding.     

Celery leaf spot: sources of inoculum

The relative importance of infected celery seed, infected leaf debris in the soil, and infected wild celery, in the incidence of Septoria leaf spot in cultivated celery has been investigated. Infection can be caused when the sole source of inoculum is viable spores on the seed surface; such spores are considered to be the main cause of disease outbreaks. Of all the seed samples examined, 93% were infected by Septoria spp. In untreated seed samples, 40% carried viable spores which survived for up to 15 months on seed stored in the laboratory, and for longer periods on seed stored at -20d? C. However, ageing of seed is not recommended as a commercial control measure. The fungus was not found in seed embryos or endosperms but mycelium was present in pericarps and testas. Unconfirmed evidence suggests that in favourable circumstances new spores might be produced in old seed-borne pycnidia.     

Colonization of Fusarium Wilt‐Resistant and Susceptible Watermelon Roots by a Green‐Fluorescent‐Protein‐tagged Isolate of Fusarium oxysporum f.sp. niveum

Interactions between watermelon and a green fluorescent protein (GFP)‐tagged isolate of Fusarium oxysporum f.sp. niveum race 1 (Fon‐1) were studied to determine the differences in infection and colonization of watermelon roots in cultivars resistant to and susceptible to Fusarium wilt. The roots of watermelon seedlings were inoculated with a conidial suspension of the GFP‐tagged isolate, and confocal laser scanning microscopy was used to visualize colonization, infection and disease development. The initial infection stages were similar in both the resistant and susceptible cultivars, but the resistant cultivar responded differentially after the pathogen had penetrated the root. The pathogen penetrated and colonized resistant watermelon roots, but further fungal advance appeared to be halted, and the fungus did not enter the taproot, suggesting that resistance is initiated postpenetration. However, the tertiary and secondary lateral roots of resistant watermelon also were colonized, although not as extensively as susceptible roots, and the hyphae had penetrated into the central cylinder of lateral roots forming a dense hyphal mat, which was followed by a subsequent collapse of the lateral roots. The initial infection zone for both the wilt‐susceptible and wilt‐resistant watermelon roots appeared to be the epidermal cells within the root hair zone, which the fungus penetrated directly after forming appressoria. Areas where secondary roots emerged and wounded root tissue also were penetrated preferentially.