UNIVERSITI TEKNOLOGI MARA INTERNATIONAL EDUCATION CENTRE (INTEC)

BIOLOGY LABORATORY REPORT A LEVEL PROGRAMME 2014

10 MARCH 2014

NAME: MAISARAH BINTI ZAINUDDIN IC NUMBER: 950201-14-5724 GROUP: 11SC6 SID NUMBER: 1311171707 TITLE: INVESTIGATING PLANT MINERAL DEFICIENCIES

1.0

TITLE
Investigating Plant Minerals Deficiencies

2.0

OBJECTIVES
To investigate the effects of different plant minerals deficiencies

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INTRODUCTION Lemna sp.i

Lemna sp. or commonly known as duckweed among society. Duckweed is the common name for a family of small aquatic herbs known as Lemnaceae. It is an aquatic plant that floats freely on or just beneath the water surface. They have the unique distinction of being the smallest flowering plants on earth which mostly do not exceeding 5mm in length. This species grow abundantly forming colonization in ponds and lakes in large number mainly by vegetative reproduction, specifically through budding. Lemna sp. is flowering plant and most of them produce sexually. Duckweed meal also high in protein, fats and fibres and its a good cattle feed.

Image 1: Budding of Lemna sp.

These plants are also unique in that they do not have any stem structures. The plant is simply a fleshy ovoid or flattened structure that may or may not bear simple roots. These highly successful plants represent the ultimate in reduction for a vascular plant. Duckweed is ubiquitous to most temperate and tropical regions of the world, making it readily available to most farmers. They are typically found floating in thick mats of

homogeneous populations in quiet streams or ponds containing high levels of organic matter. Another amazing feature of these plants is that they can double their mass in less than two days under ideal conditions of nutrient availability, sunlight and temperature. Since Lemna sp. grow rapidly in short time, it is used as a model system for studies in community ecology, basic plant biology, in ecotoxicology, in production of biopharmaceuticals, and as a source of animal feeds for agriculture and aquaculture so that no ethical issues will rise.

Micronutrients and Macronutrientsii

Sixteen elements altogether are known to be important to a plant's growth and survival. They are carbon, hydrogen, oxygen, nitrogen, phosphorous, sulphur, potassium, calcium, magnesium, copper, zinc, molybdenum, boron, iron, chloride and manganese.

Image 2: Physical Changes in Plant due to Mineral Deficienciesiii

Each of these nutrients has a critical function in plants and is required in varying amounts in plant tissue. Plants show symptoms being unhealthy when they experience insufficient nutrients. Too little or too much of any one nutrient can cause problems.

Plant nutrients fall into two categories: macronutrients and micronutrients. As the name suggest, macronutrients are those elements that are needed in relatively large amounts while micronutrients are those elements that plants needed in small amount. Both macronutrients and micronutrients are naturally obtained by the roots from the soil. Macronutrients are further divided into two groups: primary and secondary nutrients. The primary nutrients are nitrogen (N), phosphorus (P), and potassium (K). These major nutrients usually are lacking from the soil because plants use large amounts for their growth and survival. The secondary nutrients are calcium (Ca), magnesium (Mg), and sulphur (S). Micronutrients are nutrients required by humans and other organisms throughout life in small quantities to orchestrate a range of physiological functions. For people, they include dietary trace minerals in amounts generally less than 100 milligrams/day - as opposed to macronutrients which are required in larger quantities. The micronutrients or trace elements include at least iron, cobalt, chromium, copper, iodine, manganese, selenium, zinc and molybdenum. Micronutrients also include vitamins, which are organic compounds required as nutrients in tiny amounts by an organism.

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PROBLEM STATEMENT
How does absence of one or more of any macronutrients and micronutrients can lead to mineral deficiencies in plants?

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HYPOTHESIS
All plants need optimum amount of both macronutrients and micronutrients in order for them to achieve optimal growth and development. Absence of any of these macronutrients or micronutrients in plants will eventually lead to mineral deficiencies. This can be proven by observing several symptoms that are showed up in the plant.

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VARIABLES
Use different types of culture solution of same volume measured by using small beakers. Observe and record the following symptoms and

Manipulated Variable: Type of culture solution used Responding Variable:

Growth condition of the Lemna conditions of the plantlets:

sp. plantlets

a. Number of leaves b. Any physical changes such as changes in colours

Constant Variable: Type of Lemna sp. used

Use the same Lemna sp. plantlets which were taken from the same pond area and were kept in the same solution before they are being used for the experiment.

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APPARATUS
8 petri dishes with lids, small beakers, forceps and labelling stickers

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MATERIALS
Lemna sp. plantlets from pond water, distilled water, cellophane tape, 15 cm3 of different culture solutions containing: a. All nutrients b. Lack of potassium c. Lack of calcium d. Lack of phosphorus e. Lack of iron f. Lack of nitrate g. Lack of magnesium h. Lack of all nutrients (distilled water)

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TECHNIQUES
Observe and record the growth of the Lemna sp. plantlets over 10 days period which have been placed in different culture medium

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PROCEDURES
1. Eight petri dishes were rinsed with distilled water and labelled by using labelling stickers to indicate nutrient absent in each of their culture medium, as following: A B C Complete nutrients Lack of potassium Lack of calcium

D E F G H

Lack of phosphorus Lack of iron Lack of nitrate Lack of magnesium Lack of all nutrients (distilled water) Table 1: Labelling on each Petri Dishes

2. By using a small beaker, 15cm3 of the culture solution containing complete nutrients is measured and poured in petri dish labelled A. 3. Five pairs of Lemna sp. plantlets each containing two buds and roots were picked up by using a pair of forceps and gently transferred into the petri dish labelled A. 4. Steps 2 and 3 were repeated for petri dishes labelled B, C, D, E, F, G and H.

5. All of the petri dishes were then covered with cellophane tape to prevent the solution from spilling out of the petri dish. 6. The experiment was allowed to proceed for ten days and the growth of the Lemna sp. plantlets were observed and examined over the 10 days period.

11.0 RESULTS
PETRI DISH CONTENT Complete nutrients Number of Leaves All of the plantlets had one leave only One plantlet already had two Three of the plantlets had Two of them had three leaves, the other two had two leaves and the remaining One had four leaves, one had three leaves, two had two leaves and the remaining one Two had four leaves, two had three leaves and the remaining one only had one leaf FEATURES 0 2 4 DAYS 6 8 10

leaves while the two leaves rest still having one leave only while the other two had one leave only

plantlet still had only had one one leaf only Change in Physical Appearance All plantlets were green in colour Four of them were green in Three of them were green in leaf Those who had four and three leaves were green in colour while

colour while the colour while the the remaining were yellowish other one was yellowish rest looked yellowish Two of them Another plantlet was dead, two surviving The remaining three plantlets were all had two leaves

Lack of potassium

Number of Leaves

All of the plantlets had one leave only

Only one

plantlet had two had two leaves, leaves while the another two rest still had plantlets had

one leave only

one leaf only and one plantlet dead

plantlets had two leaves while the other one only had one leaf

Change in Physical Appearance

All plantlets were green in colour

One of the plantlets was transparent while the rest are yellowish green

Four plantlets looked yellowish green while one was dead

All of the surviving platelets looked yellowish and had holes on their surface

Two of them were green in colour while the other one looked pale and all of them had holes

Lack of calcium

Number of Leaves

All of the plantlets had one leaf only

Two of the plantlets started to have two

Two of the plantlets dead, one of them had

One of them had two leaves, two plantlets had one leaf only and the other two were already dead

leaves while the two leaves others remain one while the other two had one leaf only Change in Physical All plantlets were green in Three plantlets were green in Three plantlets were yellow in All of the surviving plantlets were in pale green and had holes on the surface of the leaves

Appearance

colour

colour while the colour while the other two were yellow in colour other two were in pale green.

Lack of phosphorus

Number of Leaves

All of the plantlets had one leaf only

All of the plantlets had one leaf only

Three plantlets had one leaf while the rest had two leaves.

Four of them had two leaves while the other one only had one leave

Two of them had three leaves, two had two leaves while the other one had one leaf

Three of them had three leaves, one had two leaves while another one had one leaf

Change in Physical Appearance

All plantlets were green in colour

All plantlets were green in colour

Three were green in colour while the other two were yellow in colour

Two were green Three plantlets in colour while the remaining three were in pale green were in pale green, one was green in colour while another one was yellowish

Lack of iron

Number of Leaves

All of the plantlets had one leaf only

Three plantlets had three leaves, one plantlet had two leaves while the other one was dead

Four plantlets had three leaves while the other

one was already dead Change in Physical Appearance All plantlets were green in colour One plantlet Three of them were in pale green All of the surviving plantlets were in pale green

was transparent, while the other one looked more two were in pale green and the remaining two were yellow in colour pale

Lack of nitrate

Number of Leaves Change in Physical Appearance

All of the plantlets had one leaf only

Two plantlets had two leaves, the other two plantlets had one leaf only while the other one was dead

All plantlets were green in colour

Four of them were in yellowish green while the remaining one was transparent

Two were yellow in colour with holes, the other two were in pale green with holes and the other one was dead

Lack of magnesium

Number of Leaves

All of the plantlets had one leaf only

Only one

Three plantlets had two leaves, the other two were

plantlet had two dead leaves while the

rest still had one leaf only Change in Physical Appearance All plantlets were green in colour Three of the plantlets were green in colour, one was yellow in colour while the other one was transparent Lack of all nutrients Number of Leaves All of the plantlets had one leaf only Only one Only one plantlet had three leaves while the Three plantlets were green in colour while the other two were transparent The surviving plantlets were all in very pale green

plantlet had two remaining four were all dead leaves while the rest still had one leaf only

Change in Physical Appearance

All plantlets were green in colour

Two of the plantlets were green in colour while the other started to change to yellow

Three of them were transparent, one was pale green while the other one was green

The one surviving plantlet was green in colour

The remaining plantlets looked really pale

The remaining plantlet was decreasing in size and looked really pale

Table 1: Observations on Lemna sp. plantlets over 10 days

DAY

NO OF LEAVES PRESENT 0 (dead) 1 2 3 4 Total 0 (dead) 1 2 3 4 Total 0 (dead) 1 2 3 4 Total 0 (dead) 1 2 3 4 Total 0 (dead) 1 2 3 4 Total 0 (dead) 1 2 3 4 Total

NO OF PLANTLETS HAVING PARTICULAR NO OF LEAVES IN EACH PETRI DISH A B C D E F G H 5 5 4 1 6 2 3 8 1 2 2 11 1 2 1 1 12 1 2 2 15 5 5 5 5 4 1 6 1 2 2 6 2 2 1 4 2 3 6 5 5 3 2 7 2 2 1 4 2 2 1 4 2 2 1 4 2 2 1 4 5 5 5 5 3 2 7 1 4 9 1 2 2 11 1 1 3 12 5 5 5 5 5 5 1 1 3 11 1 1 3 11 1 4 12 5 5 5 5 5 5 1 2 2 6 1 2 2 6 1 2 2 6 5 5 5 5 4 1 6 2 3 6 2 3 6 2 3 6 5 5 5 5 4 1 6 4 1 3 4 1 3 4 1 3

10

Table 2: No of Leaves of Plantlets in each Petri Dish

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TOTAL NO OF LEAVES IN EACH PETRI DISH OVER 10 DAYS

14

12 A 10 NO OF LEAVES B C

D E F

G H

0 DAY 0 DAY 2 DAY 4 DAY 6 DAY 8 DAY 10

12.0

DISCUSSION
This experiment was conducted to study the effect of various mineral deficiencies on

the Lemna sp. plantlet or also known as duckweed. Lemna sp. plantlets were used in this experiment because they are found abundantly in pond environment, easily obtained and have less ethical issues but most importantly, the effects of any deficiency of mineral can be seen quite clearly in short amount of time as this plant only possesses short life span. Amount of sunlight (by placing all petri dishes containing medium nearby window), air and temperature for the Lemna sp. plantlets were kept in control for all the cultures. This is to ensure that the only factor that will affect the growth of the Lemna sp. plantlets were the different culture solution used. As discussed in procedure, by manipulating the absence of the minerals in culture solution, the effect of mineral deficiencies that were shown physically (number of leaves and changes in colour of the leaves) by the Lemna sp. plantlets which are responding variables in this experiment, were noted. A culture medium containing all necessary nutrients were used as a control in this experiment to show the actual growth and development that should be achieved theoretically by Lemna sp. plantlets. This controlled culture medium sample was then used as comparison with other mineral deficient cultures to point out the effect resulted by specific mineral deficiency. Looking at the result , it can be concluded that presence of all the macronutrients namely nitrogen, phosphorus, potassium, magnesium, calcium and sulphur and micronutrient iron results in increase in both number of plantlets and number of green leaves. Increase in number of plantlets is due to their vegetative reproduction (budding) since there is enough availability of source and nutrients needed for their development. Average of the root length also seems to be constant and this indicates good root development despite the increase in number of the plantlets. Other that this, it also noted that only one of the green leaves turn to yellow and then white and this may due to the competition in obtaining nutrients among the plants. On the other hand, the plantlets that was grown in the solution that lack the mineral potassium had shown different growth characteristics. Out of the 5 plantlets only 3 survived over the 10 day period and only grew two leaves. Of the remaining plantlets, they all had a yellowish green colour to them and they all had holes in them. The roles of potassium in

plants are for the opening and closing of the stomata, photosynthesis and it also requires potassium ions (K+) for protein synthesis. Thus when the plantlets lacked potassium in its nutrient solution, it prevented the proper function of the opening and closing of the stomata which then prevented the entry of important gases needed for photosynthesis. Therefore this caused some of the plantlets to die off while the remaining plantlets to have slightly yellowish colours on their leaves. The plantlets that were grown in the solution that lack the mineral calcium too showed different characteristics in terms of its growth. Some of the roles of calcium in plants are that in controls the permeability of membranes and also it combines with pectin to form calcium pectate. Without calcium present in the nutrient solution, it caused the yellowing of leaves and holes which indicated death at the growing regions. The blossom-end rotting could not be seen through this experiment as this plant species that was used has no fruits. On the other hand, the plantlets that was grown in the solution that lack the mineral phosphorus had shown different growth characteristics. Phosphorus plays a major role in the growth and development of a plant. Phosphate ions are needed for the phosphate groups in adenosine diphosphate (ADP) and adenosine triphosphate (ATP), which are involved in energy transfers in cells. When phosphorus is not present in the nutrient solution or in the soil, the energy generation of a plant will be disrupted and thus, basic processes such as photosynthesis and nutrient movement cannot occur normally. Usually when a plant is deficient of the mineral phosphorus, its leaves experience dark green colouration with the purpling of veins of the older leaves as well as stunted growth. However, in this experiment the results that were obtained were quite different in which the plantlets continued to grow leaves and the colour of the leaves turned yellow instead. The plantlets that was grown in the solution that lack the mineral iron and magnesium had shown different growth characteristics as well. Both iron and magnesium are needed in the formation of chlorophyll. The condition in which leaves produce insufficient chlorophyll is known as chlorosis. As chlorophyll is responsible for the green colour of leaves, chlorotic leaves are pale, yellow, or yellow-white. The affected plant has little or no ability to manufacture carbohydrates due to the lack of chlorophyll in its leaves through photosynthesis and may die unless the cause of its chlorophyll insufficiency is treated. This is shown to be true in this experiment in which the leaves of the Lemna sp. plant turned pale yellow over the time period.

The plantlets that was grown in the solution that lack the mineral nitrogen had shown different growth characteristics as well. Nitrate ions are required to make amino acids, enzymes, and DNA which are all essential for the growth of a plant. The results obtained in this experiment had shown that the leaves of the plantlets had turned pale yellow and even had holes in them. One of the plantlet even died during the 10 day period while some experience slower growth than the other plantlets. These all coincides with the usual symptoms of nitrogen deficiency in plants. Without any essential nutrients, the number of plantlets and number of leaves goes down the slope and eventually none of the plantlet survives till the final day except for one only. This is because all the metabolic activities are restricted thus the plants cant produce any food to survive thus resulting in zero survival chance.

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FURTHER STUDIES
Another experiment can be carried out by replacing Lemna sp. plantlets with barley

seedlings using water culture technique. The seedlings are grown in culture solution (same with the above procedure) but in a test tube. The test tube should be covered by foil to exclude light, preventing algae growth. The seeds have to be moist so that it can germinate a week before use. Plants should inspect regularly for general growth, shape of leaves, length of leaf growth, colour of upper leaves, length of root growth, colour of lower leaves and mass of the seedling (before and after the experiment).iv

Image: Set up of Apparatus

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SAFETY PRECAUTION
In order to avoid any kinds of accidents or injuries during the experiment, there is a

list of precautionary steps should be taken and applied. First and foremost, wearing lab coat and a pair of closed shoes are compulsory while conducting the experiment at all times in order to protect the skin and clothing from spillage of any chemicals or substances. Besides, it is also compulsory to wash hands thoroughly with soap and water before and after conducting the experiment. This is important to avoid contamination. Furthermore, all of the glassware especially small beakers should be handled with full care since they are small and very fragile. Other apparatus such as forceps also must be sterilised before being used during conducting the experiment to prevent infection of microorganisms and must be used with full care to avoid any injury. Other than that, all of the Lemna sp. plantlets should be choose carefully as they are very fragile little creature. The petri dishes also must be rinsed with distilled water before culture medium together with the Lemna sp. plantlets being placed in them. This is to remove any impurities and microorganisms that may cause disruption to the plants growth and development. After using all of the samples and apparatus, they should be discarded properly and returned back to their original places at the end of the experiment. This is to avoid any injury and unnecessary accidents that may result fatal results.

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CONCLUSION
All plants need both macronutrients (potassium, calcium, phosphorus, nitrates and magnesium) ad micronutrients (iron) in order to achieve optimal growth and development. Deficiency or absence of any of the nutrients can be spotted through the symptoms than can be seen on its physical appearance such as changes in colours and number of roots. This proves that different types of mineral deficiency results in different effects on the growth of the Lemna sp. plantlets. Thus, the hypothesis is accepted.

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i

REFERENCES

Germplasm Resources Information Network. United States Department of Agriculture.

2006-11-03. Retrieved 2013-04-13.

ii

Flour Fortification Initiative, GAIN, Micronutrient Initiative, USAID, The World Bank,

UNICEF, Investing in the future: a united call to action on vitamin and mineral deficiencies, p. 19.
iii iv

http://hydrophytesblog.com/?paged=4. Accessed on 3th March 2012 http://www.nuffieldfoundation.org/practical-biology/investigating-effect-mineralsplant-

growth. Accessed on 3th March 2012