A new disease of cowpea caused by Alternaria cassiae

Abstract

Cowpea (Vigna unguiculata (L.) Walp is an indigenous food legume in Africa, which has great production potential, especially in areas with low agricultural resources. During surveys of cowpea fields in southern Africa, a new foliar disease was recorded. Alternaria cassiae Juriar&Khan was consistently isolated from diseased plant material. Pathogenicity was confirmed using Koch's Postulates. The effect of different culture media, temperature, light and wounding on the growth and sporulation of the fungus was studied. A. cassiae grew well and produced conidia abundantly when maintained on V8-agar at 25°C in a 12h UV-light/12h dark cycle. Sporulation was further enhanced by wounding the cultures. The pre-penetration and infection process of A. cassiae on cowpea leaves was studied by light and scanning electron microscopy. Conidia germinated within 2-3h post inoculation (hpi), forming multiple germ-tubes randomly that grew in any direction across the leaf surface. By 8hpi terminal or intercalary appressoria were formed above epidermal cells or over stomata. Occasionally germ-tubes entered stomata, without the formation of appressoria. Penetration of the plant surface, whether directly through the epidermis or indirectly through stomata was observed 72hpi. Following penetration bulbous primary hyphae were observed within the sub-stomatal cavities, secondary hyphae developed from the primary hyphae and grew within the intercellular spaces penetrating epidermis and mesophyll cells. A. cassiae is a necrotrophic fungus as the infection process is characterised by a destructive necrotrophic phase where plant cells became necrotic even prior to fungal penetration. Conidial morphology, types and development of the fungus were studied in vitro on different culture media and in vivo on cowpea leaves. A. cassiae produced a mixed population of three conidial types. Conidia were formed singly or in chains of 2-4 conidia. Conidia with long, filiform beaks and conidia with shorter beaks, converted into secondary conidiophores were more frequently produced than mature, beakless conidia on all the media, except on potato dextrose agar. Conidial body and beak sizes were variable when measured in culture and on cowpea leaves. Conidia produced in culture were larger, than those produced in vivo. Conidiophores emerged directly through the epidermis or stomata or were formed when hyphae growing on the leaf surface differentiated into conidiophores. Smooth, bud-like conidial initials were produced at the apex of conidiophores. Conidia matured and became elliptical to obvate and densely verrucose. Once a mature conidium had detached, a small pore was visible at the apex of the conidiophore. A. cassiae was shown to be seed-borne in cowpea. Six fungicides i.e. Benomyl, bitertanol, captab, mancozeb, propiconazole and triforine were evaluated for their efficacy in reducing mycelial growth of A. cassiae in vitro. All fungicides except benomyl proved to be effective. Cowpea seeds were artificially inoculated with A. cassiae and treated with all the fungicides except benomyl. Percentage germination and infection was determined in vitro. Percentage emergence, disease incidence, root and shoot lengths and abnormalities were determined in greenhouse trials. Only bitertanol at l.5x the recommended dosage significantly reduced percentage germination. All treatments except triforine l.0x and l.5x significantly decreased the percentage infection of artificially inoculated seeds. None of the treatments except bitertanol l.5x showed a difference in shoot and root length when compared to the control. Captab l.5x the recommended rate proved to be the best treatment over all.