Gaite et al: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)

Evaluation of the Presence of Kinetin in Rice (Oryza sativa)

Rafael I. Gaite1, Romeo Robert C. Indico IV2, Joshua Ivan A. Roldan3, Shannie A. Taduran (Research 2
Teacher), and Alpha Rowena O. Pimentel (Research Adviser)

Philippine Science High School – Bicol Region Campus, Department of Science and Technology,
Tagongtong, Goa, Camarines Sur, 4422, Philippines

ABSTRACT

Kinetin is a hormone found in plant and animal tissues. Studies have extracted
kinetin from plant tissues; however, these have focused on the fruit of the plant.
Rice (Oryza sativa) is a staple crop and is used as a source of food, making it an ideal
and widely available source of kinetin. This study aims to evaluate the presence of
kinetin in rice (Oryza sativa). Utilizing ethanol extraction method, samples were
extracted from different parts of the plant – fruit, roots, leaves, and stem – to
investigate the yield of different parts of the plant. To investigate the effect of a
different solvent on the yield, methanol was also used in the extraction and
compared with ethanol sample yield. The percent yield was taken; the stems
contained the highest and the roots had the lowest, for both solvents. Independent t-
Test was used due to the nature of the variables; results showed that there is no
significant difference between ethanol and methanol yields, indicating that the
solvent did not affect the yield. To evaluate the concentration of kinetin in the
sample, Ninhydrin reagent and spectrophotometry was used. All samples tested
negative for Ninhydrin reagent. The samples were tested in 268 mµ ultraviolet light,
which is the maximum absorbance point for kinetin. The roots in ethanol solvent
tested positive with maximum absorption. The results show that the roots sample in
ethanol only contained a compound similar to kinetin which may also promote
cytokinesis.

Key words: crop, evaluation, presence

Gaite, R., Indico, R.R., Roldan, J.: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)
Gaite et al: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)
INTRODUCTION
Kinetin is a plant hormone that plays a vital role
in plant growth by inhibiting chlorophyll
production and decreasing the rate of cell
division thereby effectively reducing their
growth rate. A study by Khalid et al in 2012
found that application of kinetin on kinnow
mandarins (Citrus reticulata) extended their
shelf-life (Khalid et al, 2012).
Rice (Oryza sativa) is an example of a grainy
plant and it is the most important food crop in
the world, directly feeding more people than any
other crop, according to CGIAR Research
Program on Climate Change, Agriculture and
Food Security (CCAFS). This study will
evaluate the presence of kinetin in the rice plant.
Moreover, this will assess which solvent and
plant part will produce the higher yield of
unknown compound. This study will benefit
farmers and the agriculture sector as it will bring
more focus onto their trade. Moreover, this study
will also benefit the cosmetics sector as it
employs kinetin in various aging products.
Ultimately, the study will bring more use and
importance to the rice plant and will contribute
to the study of kinetin and its effect on cell life.
METHODOLOGY
Procurement of rice plants
The rice plants were procured from a farm in
Bula, Camarines Sur wherein only 100 day-old
Gaite, R., Indico, R.R., Roldan, J.: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)
rice plants were selected for the study. At 100
days old, rice plants enter their ripening age and
kinetin levels are at their peak (Miller, 1956).
Preparation of Sample
After procurement, the rice plants were washed
to remove physical contaminants then cut up
into 4 parts: rice fruit (pre-mill rice grain),
leaves, stem, and roots. After separation, each
rice plant part was dried.
The researchers weighed 3.5 g samples of bran,
leaves and stems, and roots which were soaked
in 100mL 95% ethanol and methanol;
afterwards, the methanol and ethanol solutions
of each respective plant parts were left to stand
for 24h.
Filtration
After letting the methanol and ethanol solutions
stand for 24h, the samples were filtered using
cellulose filter papers. Afterwards, the volume
of the filtrate was measured using a graduated
cylinder.
Distillation Process
The filtrates were distilled using a simple setup
which consisted of a hot plate, condenser,
connecting glass tubes, and two flasks. The
distilling temperatures were 125° C and 155° C
for the methanol and ethanol solutions,
respectively (Miller, 1956). The concentrated
extract after distillation was measured using a
graduated cylinder.
Ninhydrin Test
 Gaite et al: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)

Test tubes were prepared for each replicate. Table 1.2 Average Percent Yield of Filtrates – 2nd set
Micropipettes were used to measure and transfer Plant Part Methanol Ethanol
the liquids. Each test tube contained 1mL of an % Yield % Yield
extract, 3 mL distilled water, and 1mL of the Fruit 29.4 25.9
Ninhydrin reagent. The test tubes were soaked in
Leaves 20.2 14.9
a water bath at 90°C for 15 min. The appearance
of the Ruhemann’s purple color serves as an Stem 17.2 13.6
indicator for the presence of kinetin (Miller,
1960). Roots 23.8 18.5

Spectrophotometry

Two (2) mL of the filtrates were transferred to

According to the first set of data, the samples
cuvettes via a pipettor. Using a UV-Vis
which contained the leaves and stems produced
spectrophotometer, the cuvettes containing the
the highest yield for both methanol and ethanol
solutions were analyzed. UV light at 268nm
solvents. However, the second set of data shows
wavelength was passed through the cuvettes and
that the fruit samples produced the highest yield
the absorbance of each sample was measured.
among the other parts. This suggests that the
The presence of kinetin will be indicated by a
plant part is not a factor in the percent yield.
maximum absorbance value at 268nm of each
Additionally, all methanol samples produced the
sample (Miller, 1968).
higher yield over the ethanol samples for all
parts. This implies that the solvent does affect

RESULTS the percent yield of the compound. However, the

percent yield does not yet indicate the presence
Percent Yield of kinetin in the samples.
Table 1.1 Average Percent Yield of Filtrates – 1st set Ninhydrin Test

Plant Part Methanol Ethanol The Ninhydrin Reagent tested negative on all

% Yield % Yield samples for both data sets. Ruhemann’s purple,

Fruit 21.07 7.96 which was not found in the samples, detects the
presence of secondary amines – a component of
Leaves and 31.22 25.4
kinetin. This indicates that the filtrates did not
Stem
contain kinetin (Miller, 1960).
Roots 7.53 4.1
Spectrophotometry

Gaite, R., Indico, R.R., Roldan, J.: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)
 Gaite et al: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)

Table 2.1 Absorbance Values of Samples in compared to the fruit and the roots. In addition,
UV-Vis spectrophotometer – 1st set the results of the t-Test show that there is no
Plant Wavelength Absorbance significant difference in the percent yield
Part (nm) between the solvents used. Spectrophotometry
Methanol Ethanol
results show that the leaf filtrates had the same
Fruit 268 0.040 0.078 absorbance as kinetin, both having a maximum
Stem at 268 nm. However, all filtrates tested negative
and 268 2.481 2.916 in the Ninhydrin test, which suggests that they
Leaves did not contain any amines, a component of
Roots 268 2.918 3.000 kinetin. This indicates that the filtrates did not
contain kinetin.
Table 2.2 Absorbance Values of Samples in
Based on the findings, the researchers conclude
UV-Vis spectrophotometer – 2nd set
that none of the filtrates contained kinetin.
Plant Wavelength Absorbance
Part (nm) For further study, the researchers recommend
Methanol Ethanol
that the cytokinin properties of rice roots be
Fruit 268 2.299 2.404 tested on plants and its growth observed, as they
Leaves 268 3.000 3.000 have similar properties to a kinetin-like
Stem 268 2.983 2.922 compound extracted by Miller in 1960, which
Roots 268 2.411 2.698 promoted cell division and prolonged the shelf
life of plants. It is also recommended that other
Absorbance is the capacity of a substance to methods of extraction for kinetin from plants be
absorb light of a specific wavelength. Based on studied, which will contribute further to the
the absorbance values, only the leaves and roots study of cytokinesis and kinetin.
filtrates have a maximum absorbance of 3.00 at
268 nm, which is the absorbance rate of kinetin
in ultraviolet light, as stated by Miller’s study in ACKNOWLEDGEMENTS

1960. Firstly, the researchers thank Mrs. Shannie

SUMMARY AND CONCLUSIONS
Based on the findings, the leaves and stems of
the rice plant (Oryza sativa) yielded the highest
percentage of the unknown compound as
Taduran for her devotion and patience towards
us and our study. Also, the researchers
acknowledge Ms. Alpha Rowena Pimentel for
her valuable insights on our study. The
researchers also thank Ms. Hannah Fenina
Estrella and Mr. Joseph Dechavez for their

Gaite, R., Indico, R.R., Roldan, J.: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)
Gaite et al: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)

critical comments on our paper. The researchers chlorophyll synthesis in tobacco tissue
also thank the SRA and SRS for their invaluable cultures. Plant physiology 47(5): 691-
help in lab and equipment acquisition and use. 695.
Additionally, the researchers would like to thank
DADZIE BK, ORCHARD JE. 1997. Routine
friends, family, for their guidance and support.
Post Harvest Screening of
Banana/Plantain Hybrids: Criteria and

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Gaite, R., Indico, R.R., Roldan, J.: Evaluation of the Presence of Kinetin in Rice (Oryza sativa)