ASSESSING PLANT SPACING AND DENSITY ON THE NUTRIENT USE

EFFICIENCY (NUE) OF NPK ON BAMBARA.

CHAPTER THREE

MATERIALS AND METHODS

3.1 Experimental Site

The experiment will be conducted at the research farm, University of Ilorin, Ilorin, Nigeria (Lat
8o 291 N, 4o 351 E), which falls within the Southern Guinea Savanna ecological zone of Nigeria.

3.2 Experimental Materials

3.2.1 Planting Materials

A single variety of Bambara groundnut (Vigna subterranea (L.) Verdc.) will be used. The seeds
were obtained from the International Institute of Tropical Agriculture (IITA) germplasm centre.

3.3 Experimental Design and Treatments

The experiment will be a 2-factor factorial arrangement laid out in a randomized complete block
design (RCBD) with three replications and twenty treatments.

The treatments will consist of:

 Plant Spacing and Density

 NPK fertilizer;

This resulted in:

3.4 Crop Establishment

Polythene bags are to be filled with topsoil, to be collected from the field (0 – 20cm). The soil
was thoroughly mixed to ensure uniformity and then filled into pots to a uniform weight of 12kg
per bag. Three seeds will be sown per pot and later thinned to one healthy seedling at two weeks
after emergence.

3.6 Crop Management

Pots will be weeded manually. No inorganic fertilizer will be applied to avoid suppressing AMF
activity. Pest control will be done using non-systemic methods when necessary.

3.8 Data Collection

I. Phenological and Early Growth Parameters

1. Days to 50% Seedling Emergence: The number of days from sowing to the date when 50% of
the planted seeds per treatment had emerged above the soil surface will be recorded through
daily observation. Emergence was defined as the visible protrusion of the cotyledon above the
soil.

2. Emergence Percentage (%): Emergence percentage will be determined at 10 days after sowing
by counting the total number of emerged seedlings per treatment and expressing it as a
percentage of the total number of seeds sown using the formula:

Number of seeds sown

3. Seedling Vigor Index: At 14 days after emergence, five seedlings per treatment will be
randomly selected. Seedling length (root + shoot) will be measured using a ruler. Seedling vigor
index was calculated as:

SVI = Emergence (%) × Mean seedling length (cm)

4. Days to First Flowering: The number of days from planting to the appearance of the first fully
opened flower on any plant within a treatment will be recorded.

5. Days to 50% Flowering: Days to 50% flowering will be recorded as the number of days from
sowing to the date when 50% of plants per treatment had at least one open flower.
6. Days to Pod Setting: Days to pod setting will be recorded as the number of days from planting
to the first visible formation of pods following fertilization.

7. Days to Physiological Maturity: Physiological maturity will be determined when pods become
hard, seed filling is completed, and leaves show yellowing and senescence. Days from sowing to
this stage will be recorded.

8. Initial Plant Stand Count: The number of established plants per treatment will be counted at
two weeks after emergence.

9. Final Plant Stand Count: Final plant stand count will be recorded at harvest to determine plant
survival under different treatments.

10. Juvenile Leaf Expansion Rate: Leaf area will be measured at two early growth stages (e.g., 3
and 5 WAS). Juvenile leaf expansion rate will be calculated as the change in leaf area over time:

LA 2 ​− LA 1
Leaf Expansion rate =
t 2−t 1

II. Vegetative and Morphological Parameters

11. Plant Height (cm): Plant height will be measured at 4, 8, and 12 weeks after sowing (WAS)
using a meter rule from the soil surface to the apical meristem.

12. Number of Trifoliate Leaves: The total number of fully expanded trifoliate leaves per tagged
plant will be counted manually at each growth stage.

13. Terminal Leaflet Length and Width: Terminal leaflets will be detached and measured using
digital vernier calipers. Length will measured from the leaflet base to the tip, while width was
measured at the widest point.

14. Petiole Length: Petiole length will be measured from the point of attachment to the stem to
the base of the lamina using a ruler.

15. Canopy Spread: Canopy spread will be measured by recording plant width along two
perpendicular directions using a measuring tape, and the mean value will be calculated.
16. Number of Primary Branches: Primary branches originating directly from the main stem will
be counted manually at peak vegetative stage.

17. Internode Length: Internode length will be determined by measuring the distance between
two successive nodes along the main stem.

18. Stem Diameter: Stem diameter will be measured 2 cm above the soil surface using a digital
caliper.

19. Growth Habit: Plants will be visually assessed to be classified as bunched, semi-bunched, or
spreading based on canopy architecture.

20. Terminal Leaflet Shape: Leaflet shape will be visually classified according to standard
descriptors (oval, lanceolate, elliptic, etc.).

21. Petiole Pigmentation: Petiole pigmentation will be assessed visually and scored based on the
presence or absence of anthocyanin coloration.

22. Root Length: At harvest, plants will be carefully uprooted, roots washed free of soil, and the
length of the longest root measured using a ruler.

23. Root-to-Shoot Ratio: Roots and shoots will be separated and oven-dried at 70 °C to constant
weight. Root-to-shoot ratio is to be calculated as:

Root Dry Weight

24. Total Leaf Area: Leaf area will be estimated using length × width × correction factor.

IV. Yield and Post-Harvest Parameters

35. Pods per Plant: Pods will be counted manually after harvest.

36. Fresh and Dry Pod Weight: Fresh pod weight will be recorded immediately after harvest.
Pods to be oven-dried at 70 °C and weighed to obtain dry pod weight.

37. Shelling Percentage: Shelling percentage will be calculated as:

Seed Weight
Shelling percentage (%) = ×100
Pod weight
38. 100- Seed weight: One hundred (100) seeds will be randomly selected from each pot and
weighed using a digital balance. The weight will be recorded in grams.

39. Seed Yield/pot: Seeds will be manually shelled from pods, then counted per pot.

40. Harvest Index:

Economic yield (seed weight)

41. Pod Length (mm): Measured using digital calipers.

42. Pod Width (mm): Measured at widest section.

43. Seed Length (mm): Measured using digital calipers.

44. Seed Width (mm): Measured at widest point.

45. Pod Texture: Visual classification (smooth or grooved).

46. Dry Pod Colour: Visual classification using colour chart.

47. Seed Coat Colour and Eye Pattern: Visual assessment.

Statistical Analysis

The data collected will be subjected to analysis of variance (ANOVA) appropriate for a factorial
experiment using statistical software. Treatment means will be separated using Duncan’s
Multiple Range Test (DMRT) or Least Significant Difference (LSD) at a 5% probability level.