26 after which the roots were rinsed three times with sterile distilled water then dried
on dry sterile blotting paper. The sterilized roots were cut into small pieces under laminar airflow and plated on PDA medium in 60 mm diameter petri dishes, from
each tomato plant root, three root segments were plated per petri dish and the petri dishes incubated for seven days under laboratory conditions. Frequency of
colonization was determined by observing the total number of roots samples per treatment where colonies of endophytic fungi emerged.
3.4 Meloidogyne incognita Egg Mass Inoculation
3.4.1 Root-knot nematode extraction and inoculation
Identification of the root-knot nematodes upto the species level forms the basic important prerequisite needed for controlling the specific species of the
nematodes. In this research the method used in the identification of root-knot nematodes was perineal pattern of the female root knot nematodes of the
Meloidogyne incognita species according to Chitwood 1949; Thorne 1961, and
also based on the morphology of the adult male root-knot nematode Einsenback Hirschmann 1981.
Root-knot nematode heavily infected tomato plants were identified from the field, the female root-knot nematodes were extracted from the roots and identified
using morphological and perineal pattern methods, the egg masses were then extracted from the roots and cultured in two week old tomato plants to act as
source of inoculum, after two months RKN egg masses were extracted from the roots and inoculated into potted tomato tomato plants already treated with
endophytic fungi. Before inoculation a rough estimate of number of juveniles from ten
sampled egg masses was made. The egg masses obtained from the inoculum source were placed in 1 NaOCl to expose RKN juveniles from the egg mass for
counting, a mean number of 350 juveniles were obtained from the 10 sampled egg masses. Three holes of 3 cm deep were made in the soil at the base of tomato
plant around the root using a 0.5 cm diameter stick, care was taken not to damage the plant roots while drilling the holes where three egg masses were inoculated at
the holes at the base of each tomato plant.
27 The plants were left unwatered until 24 hours after inoculation to enable the
juveniles that hatch from the inoculated eggs to identify the roots via the root exudates and infect the roots, after juvenile inoculation the plants were maintained
in the green house for eight weeks and watered daily, this time permitted for at least two generations of root-knot nematodes.
3.4.2 Plant management practices
Plant management practices that were carried out included, daily watering, physical removal of weeds and insects, fertilizer application N; P; K 15: 15: 15 at
the rate of 1 gplant, fertilizer application was done after every 4 weeks.
3.5 Antibiosis In vitro Test
