EXERCISE 5

BIOASSAY OF PLANT GROWTH REGULATORS1

JOSE DALE L. VIACRUCIS III

2014-36709
BOT 132 T-1L

1
A scientific paper submitted in partial fulfillment of the requirements in BOTANY/HORTICULTURE 132: Plant Growth
Laboratory under Dr. Rachel Sotto, 2nd Semester A.Y. 2017-2018 at the University of the Philippines Los Baños.
 INTRODUCTION

Plant growth substances (PGS) are naturally-occuring or synthetic low molecular weight
organic compounds that inhibit or modify a physiological response, even in extremely low
concentrations. These substances regulate cellular processes in targeted cells locally, and moved to
other locations, in other functional parts of the plant. Plant hormones or phytohormones, on the
otherhand, are naturally occuring plant growth substances. These hormones, such as auxin, cytokinin,
gibberellic acids, abscisic acid, and ethylene, determine the formation of flowers, stems, leaves,
senescence, and the development and ripening of fruit. They are vital to plant growth, and without
them, plants would mostly be a mass of undifferentiated cells (Went and Thimann, 1937).

METHODOLOGY

A. Dwarf Rice Shoot Elongation Test for Gibberellins

Dwarf rice seeds were sterilized with 1% calcium hypochlorite solution for 30 minutes and then
rinsed with water. These seeds were germinated in a petri dish. Five germinated rice seeds were placed
into test tubes which are covered with cotton plug and placed in a light bunk. These seeds were allowed
to establish in an agar medium. Into each test tubes, 1.0 ml of test solutions were pipetted in. The test
solutions were different concentrations of GA (dist. H20, 0.001, 0.01, 0.1, 1.0, 10.0, unknown) in ppm.
fter 5 days, the length of the second leaf sheath of each seedlings were measured.

B. Rice Leaf Senescence Assay for Cytokinins

Leaf sections of rice were prepared by cutting 1 cm leaf sections from the region of more or less
uniform width and then dropped directly to the test solutions. 10 leaf sections were allowed to
float in each Petri dish. The test solutions were different concentrations of kinetin (0ppm, 0.001
ppm, 0.01 ppm, 0.1 ppm, 1.0 ppm, 10.0 ppm, and one unknown). Lastly, the dishes were
incubated in the dark.

After 5 days, the chlorophyll content of the leaf sections were determined using
spectrophotometry at 645 nm and 663 nm. Chlorophyll was extracted using 80% ethanol.
 RESULTS AND DISCUSSION

Table 1. Length (cm) of second leaf sheath of dwarf rice with increasing GA3 concentration
GA3 REPLICATION
MEAN
CONCENTRATIONS I II III IV V
0 ppm 2.20 1.20 2.20 2.60 2.50 2.14
0.001 ppm 2.60 2.00 2.40 2.10 2.00 2.22
0.01 ppm 1.30 2.40 2.50 2.60 2.70 2.30
0.10 ppm 2.50 2.10 2.30 2.80 2.20 2.38
1.00 ppm 2.20 2.50 2.10 2.20 3.10 2.42
10.00 ppm 2.50 3.20 1.70 3.20 3.40 2.80

Table 1 shows the length of second leaf sheat of dwarf rice with increasing GA concentration.
As observed, there is a linear relationship between GA concentration and length of leaf sheaths. As GA
concentration increased, the length of the second leaf sheath generally increased too. Gibberellins are
known to promote leaf elongation in plants, although they have a wide range of activities including
seed germination and cell elongation (Miransari and Smith, 2009). The ability of gibberellic acid to
restore growth to dwarf mutants of pea (Pisum sativum) and maize (Zea mays), and to induce bolting in
rosette plants, prompted the suggestion that GAs might be endogenous growth regulators in higher
plants (Hedden et al., 2003).

Table 2. Absorbance values of chlorophyll extracts of rice leaf sections incubated in different
concentrations of kinetin
KINETIN ABSORBANCE VALUES
CONCENTRATIONS 645 nm 663 nm
0 ppm 0.39 0.38
0.001 ppm 0.31 0.28
0.01 ppm 0.46 0.43
0.10 ppm 0.45 0.43
1.00 ppm 0.60 0.55
10.0 ppm 0.66 0.62
 Moreover, Table 2 showed the absorbance values of chlorophyll extracts of rice leaf sections
incubated in different concentrations of kinetin. The data also showed a positive linear relationship of
kinetin concentrations and the absorbance values. The higher the concentration, the higher the
chlorophyll content and the higher the absorbance values in 645 nm and 663 nm. Cytokinins delay
senescence in plants. Addition of higher concentrations of cytokinin delayed more the senescence of
leaves. The high absorbance values obtained in high concentrations of kinetin show that chlorophyll is
still intact and not all has been degraded due to senescence.

SUMMARY AND CONCLUSION

Different bioassays of plant growth substances were made to test their certain physiological
effects on different concentrations. Gibberellins showed to promote leaf elongation in plants.
Moreover, cytokinins showed inhibitory effect to senescence. In both experiments, increasing the
concentrations also increases the effectivity of the plant growth substances to their specific actions and
activities.

REFERENCES

Alejar, A. (2001). Laboratory Manual in Plant Growth First Edition. Institute of Biological Sciences,
College of Arts and Sciences and Department of Horticulture, College of Agriculture and Food
Science, University of the Philippines Los Baños.

Miransari, M., & Smith, D. (2009). Rhizobial lipo-chitooligosaccharides and gibberellins enhance
barley (Hordeum vulgare L.) seed germination. Biotechnology, 8(2), 270-275.

Went FW, Thimann KV (1937). Phytohormones. New York: The Macmillan Company.