Chapter 3

MATERIALS AND METHODS

Location and duration of the study

The study was conducted in field experiment in an area of 360.0 m 2 located at

Brgy. Real, Monreal, Masbate from April-October 2012.

Preparation of materials

Two (2) kilograms of upland rice seeds were bought from Aroroy, Masbate. The

rice seeds were soaked overnight and incubated through rag doll method for two (2) days.

The locally-made bio-fertilizer was made in Brgy. Real, Monreal, Masbate. The

materials used in making locally-made biofertilizer were rice bran and rice hull which

were bought at the nearest rice milling station in the barangay. The rice hull was

carbonized using open type carbonizer. Five parts of chicken dung, three parts of

carbonized rice hull (CRH), and two parts of base microbial inoculants were mixed and

moistened by about 40% effective microorganisms (EM) solution. The EM solution was

made up of 3% EM-1 solution and 5% molasses. The base microbial inoculant was

formulated by anaerobically fermenting rice bran with 40% EM solution for five days.

The 12-13 cm mixture was spread and covered to allow semi-anaerobic fermentation for

five (5) to seven (7) days.

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Experimental design and treatments

The experiment was laid out in randomized complete block design (RCBD) with

six (6) treatments and three (3) replications. The experimental area was divided into three

blocks each measuring 4 x 18 meters. Each block was divided into six (6) plots

measuring 4 x 3 meters. Canals were provided between plots and blocks measuring 0.25

m between plots and 0.50 m between blocks. The experimental layout is shown in Figure

2. The study used Gi-os red rice upland rice variety which was purchased from upland

rice farmers in Aroroy, Masbate. The following were the treatments of the study:

T1 – Inorganic Fertilizer at a recommended rate and was based on the result of the

preliminary soil analysis

T2 – Locally-made bio-fertilizer which was made from mixture of carbonized rice

hull (CRH), rice bran, chicken dung, molasses and EM

T3 – Bio-N

T4 – Biogroe

T5 – Mykovam

T6 – Vital N

Inorganic fertilizer application. The application of inorganic fertilizer was based

on the initial soil analysis. Inorganic fertilizer was applied in two split application at 35

DAS and 63 DAS. The plants were fertilized with urea (46-0-0) at 12 g per plot and

superphosphate (0-20-0) at 126 g per plot.

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Figure 3. Experimental field layout

Bio-N application. Three hundred thirty three grams (333 g) seeds which were

incubated for 48 hours were inoculated using 5 g Bio-N prior to sowing.

Biogroe application. Before sowing, the plot was drenched with Biogroe water

mixture which was made from 100 g Biogroe dissolved in 2 L of water.

Mykovam application. The mykovam powder was applied at a rate of 20 g per

linear meter furrow prior to sowing.

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Vital-N application. The pre-germinated seeds were inoculated using mixture of 1

g Vital-N dissolved in 10 milliliters of water, 30 minutes prior to seeding.

Locally-made biofertilizer application. The locally-made bio-fertilizer was

incorporated into the soil one (1) week before sowing at a rate of 10 kg per plot.

Field cultural management

Land Preparation. The field was plowed with animal drawn implement and

harrowed to prepare a good seedbed and firm up the soil. The weed seeds are allowed to

germinate for two (2) weeks and followed by harrowing to destroy all germinated seeds.

Sowing. Six (6) pre-germinated Gi-os upland rice seeds were planted in furrows

with 2-3cm deep at a spacing of 10 x 25 cm.

Water Management. Only rainfall watered the Gi-os upland rice. Upland rice

purely depended on rainfall for water supply.

Pest Management. Hand weeding was done 30-40 days after sowing (DAS). This

was done to prevent weed and rice competition for nutrients and light interception that

would drastically reduce grain yield. Natural control for insect pests was done.

Harvest Management. Harvesting was done when 80-85% of rice grains in the

panicle are fully ripened or when the moisture content of the grains was 20-24%. The rice

plant was cut from the base for biomass determination using sickle. The harvested rice

was threshed manually, cleaned, sun dried to about 13-14% moisture content and

weighed.
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Data Gathered

1. Agronomic Characteristics

a. Plant height. Plant height was measured from ground level to the tip of longest leaf of

rice plants from five hills which were randomly selected. Plant height measurement was

done at weekly interval starting at 21 DAS.

b. Leaf Area Index (LAI). Leaf Area Index (LAI) was determined at 42, 62, 82 DAS, and

102 DAS. Plant leaves in one hill were measured. The leaf area was calculated using

simple formula based on leaf length (LL), maximum width (LW) and correction factor (k)

for rice (k=0.75). The correction factor (k) value used at seedling and maturity was 0.67.

The area occupied per hill was 25 cm2. The leaf area index was calculated using the

formula:

∑ Leafarea x 0.75( c m 2 )
LAI =
Land area covered per hill(cm 2)

2. Physiological characteristics of upland rice

The physiological characteristics of upland rice were determined at 62, 82 and

102 DAS. The data required in the computation for each physiological characteristic were

gathered.
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a. Crop growth rate (CGR)

CGR=(W 2−W 1)/¿

Where:

CGR – crop growth rate (g/m2/day)

W1& W2 – dry weight at the beginning and end of the intervals (g)

GA – land area occupied by plants at each sampling (m2)

t1 & t2 – corresponding days (day)

b. Relative growth rate (RGR)

RGR= ( W1 ) X ( dW
dT )

RGR= ( ¿ WT 2−¿ W1
2−T 1 )

Where:

RGR – relative growth rate (g.g-1dry weight/day)

W – dry weight (g)

dW/dT – change in dry weight per unit time (g dry weight/day)

W1& W2 – dry weight at the beginning and end of the intervals (g)

GA – land area occupied by plants at each sampling (m2)

T1 & T2 – corresponding days (day)

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c. Net assimilation rate (NAR)

NAR= ( 1A ) X ( dW
dT )

Where:

NAR – net assimilation rate (g/m2 leaf area/day)

A – leaf area (m2)

dW/dT – change in plant dry matter (g/day)

d. Plant dry matter production

Plant samples were gathered from one (1) hill per plot. Initial weight of each plant

samples was determined prior to oven drying for 72 hours.

3. Plant and Soil Laboratory Analysis

The plant and soil samples were sent to Regional Soils Laboratory (RSL) at

Naga City for the following laboratory analysis:

a. Plant Tissue analysis. Plant tissue analysis was done at flowering stage. Leaf samples

were collected for nitrogen analysis.

b. Soil analysis. Soil analyses were done before sowing and after harvesting. Soil pH, %

organic matter (OM), % total nitrogen, % base saturation (BS), cation exchange capacity

(CEC) and bulk density were determined. The soil temperature, moisture content and pH

were monitored weekly using 3-in-1 soil test meter

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4. Yield and Yield Components

a. Number of tillers. The number of tillers was gathered from five (5) hills which were

randomly selected per treatment or plot.

b. Number of panicles. The number of panicles was gathered from five (5) hills which

were randomly selected per treatment or plot and was converted to number of panicles

per square meter.

c. Percent productive tillers. The percent productive tiller was determined by dividing the

number of panicles produced by the highest number of tillers produced multiplied by 100.

d. Total number of spikelets per panicle. The total number of spikelets was determined

from five (5) hills which were randomly selected per treatment or plot. Total number of

spikelets per panicle was counted from 10 randomly selected panicles.

e. Number of filled spikelets per panicle. The number of filled spikelets per panicle was

taken from ten randomly selected well-developed panicles at harvest.

f. Percent filled spikelets. The percent filled spikelets were determined by dividing the

total number of filled spikelets by the total number of spikelets multiplied by 100.

g. 1000-seed weight. The weight of 1000 seeds was taken at 13% moisture content.

h. Grain yield. Harvesting was done when 80-85% of the grains in the panicle had turned

yellow. The grain yield was gathered from 4 m 2 harvestable areas in each experimental

plot, threshed, cleaned and weighed. The moisture content (MC) of each sample was

taken using digital moisture meter. Grain yield was computed as follows:
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10 xWx(100−MC)
Yield at 13 % MC (t /h a)=
A x (100−13)

Where:

W = initial weight (kg)

MC= moisture content (%)

A = harvestable area (m2)

i. Straw weight. The rice straw from 4 m2 harvestable area in each experimental plot was

weighed.

j. Harvest index. The harvest index was computed by dividing the economic yield or

grain yield by biological yield. Biological yield refers to sum of straw weight and grain

weight.

5. Cost and return analysis

The profitability of upland rice production using different bio-fertilizers was

determined by simple cost and return analysis.

6. Statistical analysis used

The analysis of variance (ANOVA) for Randomized Complete Block Design

(RCBD) was used to determine the differences among treatments in terms of the different

parameters. The means of the different treatments were compared using the Duncan’s
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Multiple Range Test (DMRT). The soil and plant tissue analysis were quantitatively and

qualitatively described.