Funded by GCGRA and University of Ghana/Govt of Ghana
(1991 to 1991)
Researcher: S. P-K Entsie (Supervisor: Dr. G.T. Odamtten)
Samuel Paa-Kwesi Entsie (1992) MPhil Thesis, University of Ghana
Summary:
The fungus Aspergillus niger to be used in attacking the saprophytic phase of Phytophthora palmivora in soil and in stem as canker was present in the soil of the selected cocoa farms at Suhum, Kade and Nankese (Eastern Region, Breman (Central Region) anj Fume (Volta Region) throughout the year.
Total population of soil fungi resident in the top (0-5cm depth) layer was generally 2-5 per cent higher than what existed at the lower (5-lOcm depth) level. Total fungal population was generally low (1.9 3.8 log10CFU/g) from December to April during the dry season and thereafter increased reaching peak value (4.0-4.6 log10CFU/g) in May-July in the major rainy season and September-October in the minor rainy season.
Population of A.niger in the soil from the selected cocoa farms differed considerably throughout the sampling period; the farms at Suhum, Kade, Nankese, Breman and Fume showed unique phenology of A.niger propagules. The pH of the soil also varied as follows throughout the year: Suhum (pH 6.5-7.8), Kade (pH 5.6- 6.8), Nankese (pH 6.8-8.0), Breman (pH 5.8-6.7), Fume (pH 6.8-7.8). The top 0-5cm depth recorded pH's that were 2-4 percent higher than the lower 5-10cm depth of soil. The moisture content range of soils from the selected farms are as follows:Suhum (12.4-32.0 per cent), Kade (12.1-22.3 per cent), Nankese 2?-26.8 per cent , Bremen (6.5 -21.7%) and Fume (8.6- 23.2 per cent). Moisture content of top soil (0-5 cm depth) was 2-8% lower than what existed in deeper (5-10cm depth) layers.
In order to augment A niger populations in cocoa farm soil, three substrates were tested for their ability to support rapid growth of A niger. The best growth of A niger at 35’C was obtained on blended dry cocoa leaves. The fungus covered the entire petri plate (12.5cm diameter) in about 5 days; it took 21 days to cover the same plate containing blended dry pawpaw leaves and dry plantain leaves was clearly unstuiable as the fungus grew sparingly and took 3 months (90 days) to cover the petri plate.
Germination and growth of cocoa seedlings sown in pots containing unsterilized cocoa farm soil amended with A niger (raised on blended dry cocoa leaves) was markedly similar to seedlings raised in A niger-free unsterilized cocoa farm soil. There was no significant difference (P =0.05) in the height, dry weight of stem. root and leaves of all the seedlings after 8 weeks growth in their respective soils. Furthermore, ? seeds of 20 plants belonging to 15 Families and 21 genera germinated and developed to maturity in both non-sterile control soil and in soil amended with A.niger. A niger failed to produce in native soil the characteristic metabolite which is known to depress height and growth of cocoa and other seedlings.
Culture filtrate of Asperfillus niger required for studies on the effect of metabolite of the fungus on pre and post-emergence damping off of cocoa seedlings as well as stem canker caused by P. palmivora was provided by cultures grown in V-8 Broth of pH 2.3 at 35’C for 4 days.
A niger culture filtrate protected Amazonia, Amelonado variety, Clones T79/501,PA7/808 and NA32 from severe damping-off when beans were placed for 30 min. 1:1-1:2 v/v dilution of filtrate in the presence of P. palmivora zoospores prior to sowing in soil. There was varying levels of resistance/susceptibility to P.palmivora infection depending on the variety/clone used. Filtrate dilution of 1:1 v/v depressed height of seedlings by about 4.5 per cent.
When the beans were iwuersed in 1:1 1:2 v/v dilution of A.niger culture filtrate for·30min, 60mln and 120min before artificial inoculation with zoospores of P.palmivora, 90 per cent of T79/501 seedlings survived pre-and post emergence damping off after 10 weeks growth in soil; 80 per cent of Pa7/808 and 50 per cent of Na32 clone survived during the same period of growth. Control beans soaked in distilled water prior to artificial inoculation with P.palmivora zoospores virtually failed to germinate.
There was no statistical difference (P < 0.05) between results obtained after 30min, 60min and 120 min soaking in 1:1 v/v dilution of A.niger.
External application of culture filtrate of A.niger (1:1 - 1:2 v/v dilution) to soil variably depressed canker formation and symptoms arising from it in cocoa seedlings of crosses T63/971 x T60/887; T65/971 x AMEL; T85/799 x Pa7/808; T85/799 x T65/283 and ALPH B63xS84. Cross T85/799 x Pa7/808 was the most resistant variety followed by T85/799 x T65/283. The cross between ALPH B63 x S84 was the most susceptible to stem canker infection.
Other studies reported in this thesis show that P. palmivora could be isolated from soil throughout the year on the selected cocoa farms. The inoculum was 11.4-24.1 per cent higher in the top soil (0-5cm depth) than in the bottom soil (5-lOcm depth). The farmers did not adhere to the cultural practice of burning infected pods on the farm. This resulted in a high percentage (58-82 per cent) of farms acting as reservoir of the pathogen. The nutrient composition of the soil (Organic matter Carbon content, inorganic elements available Nitrogen and Phosphorous etc) and the prevailing microclimatic conditions on the farms presumably acted in concert to augment epiphytotic induction of the disease.