Seed Germination

Seed Germination

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Introduction:
Seed germination is a process where the quiescence seed imbibed with water, the seed radical elongates and breaks through the seed coat surrounding the embryo. However, most of the plant seeds do not germinate immediately and they under goes period of dormancy. These dormant seeds are normally dehydrated, containing inactive enzymes and resistant to unfavorable condition. These characteristic allowed the seed to be dispersed before germination occur and survive adverse environmental condition such as drought.
In order for a seed to break its dormancy, optimum conditions were required for seed to germinate. There are three important conditions for seeds to germinate: suitable temperature, sufficient water supply, and also satisfactory level of oxygen. Water supply is important as it is used to rehydrate the seed tissues and induce breakage of the seed coat which posted as a physical barrier for radical to elongate (Debeaujon et al., 2000). Besides that, oxygen is also an essential element for seed germination because aerobic respiration of the seed tissues produces energy, ATP, which is required for further development of embryo. The optimum temperature for potential seed germination varies on plant species but it is one of the important factors that affect seed germination. Factors such as light and hormones also contribute in seed germination process. The exposure of light is critical for certain plant seeds to germinate, furthermore, light have an indirect effect to the surrounding temperature. Hormones are important as they signal the production of enzyme in seed to perform process such as mitotic division, degradation of cell wall, and cell differentiation.
Thus, the aim of this experiment is to investigate the effect of light, planting depth and hormones on seed germination.





Materials and Methods:
In section A, effect of light on seed germination was studied. The materials used in section A was 100 lettuce seeds, 100 green beans, 4 Petri dishes, cotton wool, and 4 filter papers. 2 Petri dishes were labeled as water + light (LS) and water + dark (LS). The other 2 Petri dishes were labeled as water + light (GB) and water + dark (GB). All the Petri dishes were placed with filter paper that was moistened by 2mL of water. Then, 5mL of water is used to moisten the cotton wool which was placed on top of all the Petri dishes. 50 lettuces seeds were placed to each Petri dish that was labeled LS and 50 green beans were placed to each Petri dish that was labeled GB.

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The lid of each Petri dish was closed and kept at designated light or dark area.
In section B, effect of planting depth on seed germination was studied. The materials used in section B were 40 lettuce seeds (LS), Green beans (GB), and 4 pots containing soil. All pots were labeled as “bean-deep”, “bean-light”, “lettuce-deep”, and “lettuce-light”. 20 green bean seed were gently pressed into the soil of “bean-deep” pot for 1.5-2cm. The implanted beans were covered with soil and water was sprinkled to moist soil. Another 20 green bean seed were placed on the surface of “bean-light” pot and water was sprinkled to moist the soil. These steps carried out in section B were repeated for lettuce seeds by planting into “lettuce-deep” and “lettuce-light” pots.
In section C, effect of hormones inhibitor on seed germination was studied. The materials used in section C were 150 lettuce seeds, 5mL of tomato juice, 5mL of 1mM abscisic acid, 3 Petri dishes, and 3 filter papers. 3 Petri dishes were labeled as “water + light”, “tomato juice + light”, and “abscisic acid + light”. Filter paper was placed to all Petri dish. 5mL of distilled water, tomato juice, and abscisic acid were respectively poured into the labeled Petri dishes. 50 lettuce seeds were placed into each Petri dish. The lid of these Petri dishes were closed and kept under the light.
The numbers of seeds germinated for all section were recorded on the 8th days which was expressed as percent of germinated, Germination (%) =(number germinated)/50 ×100% . All the recorded data and observational data were tabulated in results.

Results:
Section A: Effect of light on seed germination
This section of experiment is carried out to study the effect of light on seed germination where both green beans and lettuce seeds were planted under both light and dark condition. The number of seed germinated for both conditions on the 8th day was recorded and expressed as percentage of germinated. The characteristic of the germinated seed were also observed and recorded in table 1.
Table 1. The number of seed germinated, percentage of seed germinated, mean percentage of seed germinated, and characteristic of the germinated seed under different light condition for both lettuce seeds and green beans on the 8th day.
Type of seeds Conditions Total seeds planted Number of seed germinated Percentage of seed germinated (%) Observation
Green bean Water and light 50 50 100 Green and larger leaf
Water and dark (absence of light) 50 49 98 Yellowish and smaller leaf
Lettuces Water and light 50 27 54 Green and larger leaf
Water and dark (absence of light) 50 48 96 Yellowish and smaller leaf

The results in section A showed that green bean germinate better with the presence of light and lettuce seed germinate better with absent of light. Based on Table 1, green beans had percentage of germinated seed of 100% and 98% for present and absent of light respectively. The green bean that germinated in a light condition had green and larger leaf compared to green bean that germinated in dark condition which had yellowish and smaller leaf. Hence, green beans had a greater germination percentage, 100%, and had a favorable physical characteristic, green and larger leaf, in the present of light compared to in the dark condition which had 98% of germinated seeds that had yellowish and smaller leaf. Table 1 also showed that the lettuce seeds had 54% of germinated seed on the 8th day in the light condition. In the dark condition, lettuce seed had 96% of germinated seed. Lettuce seed had green and larger leaf in light condition and yellowish and smaller leaf in dark condition as their physical characteristic. Therefor, lettuce had a greater percentage germinated seed, 96%, but had less favorable physical characteristic which are small yellowish leaf in the dark condition as compared to in the light condition which had 54% of germinated seed with large green leaf.

Section B: Effect of planting depth on seed germination
This section of experiment is carried out to study the effect of planting depth on seed germination where both green beans and lettuce seeds were planted in both shallow and deep planting depth. Deep planting depth is represented as planting 1.5-2cm from the soil and shallow planting depth is represented as planting on the surface of the soil. The number of seed germinated for both conditions on the 8th day was recorded and expressed as percentage of germinated. The characteristic of the germinated seed were also observed and recorded in table 2.





Table 2. The number of seed germinated, percentage of seed germinated, mean percentage of seed germinated, and characteristic of the germinated seed under different soil depth condition for both lettuce seeds and green beans on the 8th day.
Type of seeds Conditions Total seeds planted Number of seed germinated Percentage of seed germinated (%) Observation
Green bean Light (shallow) 20 5 25 Green leaf
deep 20 13 65 Green leaf
Lettuces Light (shallow) 20 6 30 Green leaf
deep 20 9 45 Green leaf

The results in section B shows that both green bean and lettuce germinate better in deep planting depth. Based on table 2, green beans had percentage seed germinated of 25% on the 8th day in the shallow soil planting depth condition. While, the seed germinated percentage for deep soil planting depth condition were 65% on the 8th day. Thus, green bean germinate better in deep planting depth, 65% of germinated seed, compared to light planting depth, 25% of germinated seed. The lettuce seeds have percentage of seed germinated of 30% on the 8th day in the shallow soil planting depth condition. While, the percentage seed germinated for deep soil planting depth condition were 45% on the 8th day. Both lettuce seeds and green beans had the same physical observation which was having green leaf. Lettuces seeds germinate better in deep planting depth, 45% of germinated seed, compared to light planting depth which is 30% of germinated seed.



Section C: Effect of hormone inhibitor on seed germination
This section of experiment is carried out to study the effect of hormones inhibitor on seed germination where lettuce seeds was planted using abscisic acid, tomato juice and water. The number of seed germinated for all listed conditions on the 8th day was recorded and expressed as percentage of germinated. The characteristic of the germinated seed were also observed and recorded in table 3.
Table 3. The number of seed germinated, percentage of seed germinated, mean percentage of seed germinated, and characteristic of the germinated seed under tomato juice treatment, abscisic acid treatment and water treatment for lettuce seeds on the 8th day.
Type of seeds Conditions
(Treatment) Total seeds planted Number of seed germinated Percentage of seed germinated (%) Observation
Lettuce Tomato juice 50 0 0 Full of fungi/mold
Abscisic acid 50 17 34 Green leaf
Water 50 22 44 Green leaf

The result of this section showed that tomato juice has completely inhibits seed germination. While, abscisic acid was better at inhibiting seed germinate compared to water. Based on table 3, there were no seed germinate under tomato juice treatment on the 8th day. The Petri dish used in this treatment had been overgrown by fungi/mold. The Petri dish that used abscisic acid treatment had the percentage of germinated seed of 34% on the 8th day. While, the petri dish that used water treatment had the percentage of germinated seed of 44% on 8th day. Both abscisic acid and water treatment had the same physical observation where both seedlings had green leaf.



Discussion:
An investigation for effect of light on seed germination were carried out in section A. Based on the result obtained in section A, green bean had higher percentage of seed germinated in light condition and lettuce seed had higher percentage of seed germinated in dark condition on the 8th day. The results obtained do not support the theoretical hypothesis which proved that some variety of lettuce seed such as Lactuca sativa seed did not germinate in dark condition (Black et al., 2006). However, lettuce seed contain phytochrome which is a photoreceptor that takes part in seed coat degradation and response to red and far red wavelength (Bae and Choi, 2008). If the seed is exposed with red light, the seed coat will degrade and hence germination occurs. Therefor, the result obtained for lettuce seed in this section might be caused by sufficient exposure of red light which was present in background lighting before incubating into the drawer. The result for green bean did not have distinct different in the percentage of seed germinated in both light and dark condition. Hence, germination of green bean does not affected by light. However, the physical appearance for the green bean that germinated in dark is similar to germinated lettuce seed in dark condition. Hence, green bean could contain phytochromes which are more sensitive thus resulted similar percentage of seed germinated in both light and dark conditions.
In section B, the effect of planting depth on seed germination was studied. The results showed that both green bean and lettuce plant had a higher percentage of germinated seed in deep planting depth compared to light planting depth. According to Harper and Benton (1966), burial at shallow depth promote more seeds to germinate compared to seeds that are lying on the surface of soil because the covered soil maintain the moist environment around the seeds. The moist surrounding induces seed to carry out imbibitions process which resulting rupture of the seed coat and elongation of radical. Therefor, the results obtained in section B fits the theoretical hypothesis which both green bean and lettuce seed that were buried under 1.5-2cm soil had the higher percentage of germinated seeds compared to the shallow-planting depth.
The effect of hormones on seed germination was investigated in section C. Based on the result obtained, tomato juice had completely inhibit the germination of lettuce seed, while, abscisic acid had a lower percentage of germinated seed compared to control, water treatment. According to Eiji et al. (2010), abscisic acid is a hormone that regulates seeds germination and also contributes to its dormancy. Abscisic acid inhibit the formation of cell wall degrading enzyme hence prevent the breakage of seed coat (Müller et al., 2006). Thus, the percentage of germinated seed in abscisic acid treatment was lower compared to water treatment. Tomato juice contains inhibitor such as ferulic acid (Sharma, 2013). High concentration of ferulic acid can inhibit seed germination (Li et al., 1993). In this experiment, the result proved that ferulic acid did inhibit seed germination. The water treatment had the highest percentage of germinated seed because the hormones in the seed were not manipulated and hence the water treatment represents the control condition.
There are a few limitations present in this experiment. One of the limitations is too little sample used as a representative for the plant species population. This limitation may lead to result that may be conflicting with the population result. Therefor, a larger sample size should be use in the future experiment. Controlled factor in the experiment should be kept constant at all times. In this experiment, over exposure of light to the seed before incubation may produce undesirable results. Thus, placing of these seed should be done at constant speed.

Conclusion:
This experiment investigate on the effect of light, planting depth, and hormones on seed germination by examining the percentage of germinated seeds based on the number of germinated seed on the 8th day. Results in section A showed that green bean had higher percentage of germinated seed with the presence of light and lettuce had higher percentage of germinated seed with absent of light. In section B, both green bean and lettuce seed had higher percentage of germinated seed in deep planting depth compared to light planting depth. In section C, tomato juice treatment had lowest percentage of germinated seed, while, abscisic acid had lower percentage of germinated seed compared to water treatment. Based on the stated information, a higher yield and efficient plant cultivating process for both lettuce and green bean can be produced which also increase the profit by the agricultural sector.

References:
Bae, G. and Choi, G. 2008, ‘Decoding of light signals by plant hormones and their interacting proteins’, Annual Review of Plant Biology, No. 59, pp.286-310.

Black, M., Bewley, J. D., Halmer, P. 2006, ‘Encyclopedia of seeds’, Science, technology and uses CAB International, Oxfordshire, pp. 376-378.

Debeaujon, I. and Koornneef, M. 2000, ‘Gibberellin requirement for Arabidopsis seed germination is determined by both testa characteristics and embryonic abscisic acid’, Plant Physiology, No. 122, pp.420–423.

Eiji Nambara, Masanori Okamoto, Kiyoshi Tatematsu, Ryoichi Yano, Mitsunori Seo and Yuji Kamiya 2010, ‘Abscisic acid and the control of seed dormancy and germination’, Seed Science Research, No.20, pp 57-60.

Harper, J. L. and Benton, R. A. 1966, ‘The germination of seeds on the surface of a water supplying substrate’, The behavior of seeds in soil, pp.153-155.

Li, H. H., Masafumi Inoue, Hiroyuki Nishimura, Junya Mizutani and Eiji Tsuzuki 1993, ‘INTERACTIONS OF trans-CINNAMIC ACID, ITS RELATED PHENOLIC ALLELOCHEMICALS, AND ABSCISIC ACID IN SEEDLING GROWTH AND SEED GERMINATION OF LETTUCE’, Journals of Chemical Ecology, vol. 19, No. 18, pp. 1778-1780.

Muller, K., Tintelnot, S. and Leubner- Metzger, G. 2006, ‘Endosperm- limited Brassicacae seed germination: abscisic acid inhibits embryo- induced endosperm weakening of Lepidium sativum (cress) and endosperm rupture of cress and Arabidopsis thaliana’, Plant Cell Physiology, No. 47, pp. 864- 870.


Sharma, J. P. 2013, ‘Plant Growth and Movement’, Comprehensive Biology XII, pp. 618.

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