BIOLOGY LAB REPORT

TITLE

: INVESTIGATING EFFECT OF PLANT MINERAL DEFICIENCIES

PREPARED BY

I/C NUMBER

STUDENT ID

GROUP

LAB PARTNER

LECTURERS NAME

PRACTICAL DATE

SUBMISSION DATE

Abstract
Plants suffering from lacking of mineral grow poorly and manifest various signs of the deficiency
in their physical appearance. In this experiment, we will grow Lemna sp. Plantlets under
controlled conditions. Following range of nutrient solutions to explore were chosen : nitrogen,
phosphorus, magnesium, potassium, calcium, iron and sulphate. The plantlets were then placed
in a medium that were lacking in selected nutrient and also two more of plantlet sets were
placed in fully absent and present of all-nutrient medium. They were then, observer for 14
constituent days for symptoms and recorded.
INTRODUCTION
1. Lemna sp. (1)
Lemna sp. or commonly known as duckweed among society is an aquatic plant that float freely
on or just beneath the water surface. Most are small, 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. are flowering plant and most of them
produce sexually. Duckweed meal also high in protein, fats and fibres and its a good cattle feed.

Figure 1 : Lemna sp. (2)

Figure 2 : Budding of Lemna sp. (3)

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.

When Lemna sp. invades a waterway, it can be removed mechanically, by the addition of
herbivorous fish or treated with a herbicide. The rapid growth of duckweeds finds application
in bioremediation of polluted waters and as test organisms for environmental studies. It is also
being used as an expression system for economical production of complex biopharmaceuticals.

2. Chlorosis (4)
Chlorosis is a condition in which leaves become pale, yellow or yellow-white due to insufficient
production of chlorophyll. The affected leaves will unable to perform photosynthesis thus,
unable to manufacture carbohydrate which will lead death of the plant due to insufficient
nutrients. Chlorosis may caused by a specific mineral such as iron, nitrogen and magnesium
deficiency in the soil, poor drainage damaged or compacted root exposure to sulphur chloride
and many more.

Figure 3 : Chlorosis of leaf (5)

3. Nutrients (6)

Sixteen elements 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,
molyhdenum, boron, iron, chloride and manganese.

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
2 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 sulfur (S).

Figure 4 : Essential Plant Nutrients(7)

Figure 5 : Image of plant that experience deficiency of nutrient(8)

Table 1 : Some of the nutrients physiological role and deficient symptom (9)

Objective
To investigate the effect of plant mineral deficiencies.
Problem Statement
How do different mineral deficiencies affect the growth of the Lemna sp. plantlets?
Hypothesis
Plants need both macronutrients and micronutrients in right proportion to achieve optimal
growth and development. Absence of one or more of these nutrients can lead to mineral
deficiencies in plants and these mineral deficiencies can be found out using several symptoms
that are showed up in the plant.

Variable :
Types of Variables

Ways to control the variables

Manipulated Variable:
Type of culture solution used

Use different type of culture solution of same

amount measured using small beakers.

Responding Variables:
Growth condition of the Lemna sp. Plantlets

All the symptoms and conditions (number of

plantlets, number of dead and green leaves, and
root length) were observed, measured and
noted.

Fixed Variables:
Type of Lemna sp. used

All the Lemna sp. Plantlets were taken from

same pond area and were kept in same
solution in laboratory.

Number of leaves in on Lemna sp. plantlet

Lemna sp. With 2 leaves (that were almost

identical in colour and size) were used
throughout the experiment.

Volume of culture solution used (ml)

Use small beakers to measure 15ml of each

nutrient solutions before the Lemna sp. Is
cultured.

Apparatus
9 petri dishes with lids, small beakers, forceps, labeling stickers, cling film, paper, black pen.
Materials
Lemna sp. plantlets (from pond water), distilled water, a range of culture solutions (15cm 3)
containing :

All nutrients

Lack of nitrogen

Lack of phosphorus

Lack of potassium

Lack of magnesium

Lack of calcium

Lack of iron

Lack of sulphur

Lack of all nutrients

Procedure
1. Nine petri dishes were rinsed with distilled water and labeled with stickers indicating
nutrient absent in each of their culture medium, as following:

All nutrients

Lack of nitrogen

Lack of phosphorus

Lack of potassium

Lack of magnesium

Lack of calcium

Lack of iron

Lack of sulphur

Lack of all nutrients

2. Using a small beaker, 15cm3 of the culture solution containing all nutrients (nitrogen,
phosphorus, potassium, magnesium, calcium, iron, sulphur) is measured using a small beaker
and was poured into its respective petri dish.
3. Five pair of Lemna sp. each containing two buds and roots was picked up using a pair of
forceps and gently transferred into the respective petri dish.
4. Steps two and three were repeated for other petri dishes.
5. The petri dishes were then covered with cling film was used to prevent the solution from
dripping out of the petri dish.
6. After all the petri dishes were placed in the same brightly lit area, the cling film was removed
gently.

7. The experiment was allowed to proceed for fourteen days and the growth of Lemna sp.
plantlets was observed and examined every day with any changes observed noted.
8. The growth of Lemna sp. plantlets were measured using a scale that was drawn on a paper
using black ink pen. Since the petri dish is transparent thus, it was placed above the scale
paper and adjusted to obtain the reading.
9. The data was collected were based on the number of platelet, the number of green leaves, the
number of dead leaves (that are white or yellow), and the average root length.

Safety precaution
In order to avoid any accident or injury during the experiment in laboratory, the precautionary
steps should be taken and applied.

Wearing lab coat and a pair of suitable shoes are

compulsory when conducting an experiment in the lab at all times to protect the skin and
clothing from spillage of any substance. Washing hands thoroughly with soap and water
before and after conducting experiment is vital to avoid contamination. Furthermore, the
glassware such as small beakers should be handled with full care because they are fragile. The
apparatus such as forceps is also sterilized to prevent infection of microorganism and should
be used with care to avoid any injury. After using all samples and apparatus at the end of
experiment, they should be discarded properly and returned back to their places to avoid
injuries and unnecessary accidents that may result fatal results.

Risk Assessment
The Lemna sp. plant should be choose carefully as they are very fragile little creature. Other
than that, once apparatus (such as small beakers) was used, they were washed and placed in
their place. The petri dishes were rinsed with distilled water to remove any impurities and
microorganism that may cause disruption to the plants health and growth.

Results

Solution
All
Nutrients
Present

Observations

Number of
Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)
Lack of
Number of
Nitrogen
Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)
Lack of
Number of
Phosphorus Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)
Lack of
Number of
Potassium
Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)

Days
0
5

2
5

4
6

6
6

8
7

10
8

12
10

14
13

10

10

14

15

15

18

19

19

1Y

1W

1.70

1.80

1.58

1.60

1.65

1.52

1.50

1.49

10

10

10

2Y

2Y,
1W

3Y,
2W

2Y,
2W

2W

0.76

0.76

0.72

0.71

0.61

0.62

0.59

0.55

10

11

11

10

1Y

1W

2W

2W

1.20

1.25

1.48

1.04

1.01

1.25

1.18

0.99

10

11

11

12

10

1Y

1Y

2W

1Y

2Y

2Y

1.04

1.04

1.33

1.35

1.36

1.24

1.25

0.78

Lack of
Number of
Magnesium Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)
Lack of
Number of
Calcium
Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)
Lack of
Number of
Iron
Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)
Lack of
Number of
Sulphur
Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)
Lack of all
Number of
nutrients
Plantlets
Number of
Green Leaves
Number of
Dead Leaves
(Yellow/White)
Average Root
Length (cm)

10

10

1Y

1Y

1W

2Y

2Y

1Y,
2W

1.32

1.20

1.25

1.54

1.63

1.15

1.24

0.99

10

10

2Y

2Y

4W

4W

3W

3W

1.30

1.40

1.33

0.87

0.73

0.69

0.70

0.53

10

10

12

14

14

16

17

17

1Y

1W

1.16

1.26

1.18

1.23

1.05

1.32

1.05

1.34

11

11

10

10

11

13

14

17

18

20

1W

1.88

1.38

1.24

1.23

1.15

1.30

1.14

1.53

10

10

2Y

2Y,
1W

3W

2Y,
1W

2W

3Y

1.30

1.08

1.20

1.23

0.82

0.63

0.54

0.32

Data Interpretation(10)
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 plantlets, number of green
leaves, number of dead leaves that are yellow or white or being in different state or colour and
average root length) 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.

Control Medium
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.

Figure 1 : Some of the Lemna sp. plantlets in control medium at the end of the observation
day.

From the results that were obtained, it was shown that the Lemna sp. plantlets were affected in
several ways due to the following mineral deficiencies :

Without Nitrogen
From Table 1, it can be seen that the number plantlets stays the same for six days before
it reduces into four and three by the end of observation period. Number of green leaves
also noticeably reduce although it remained constant for four days. Number of dead
leaves (9 yellows and 7 white leaves) in this medium is also at high rate and the average
length of root linearly decreases with observation days. These may be due to inability of
the plantlet to synthesis proteins, nucleic acids, chlorophyll and enzymes for
photosynthesis and respiration continuously thus, halting the leaf growth and root
development. The plantlets also seemed to be having stunted growth as no distinct
differences can be seen in its shoot development. The green leaves also become paler
until the end of the observation day none of the leaves were in actual green colour. Thus,
it is concluded that disruption in chlorophyll production lead to chlorosis.

Figure 2 : Some of the Lemna sp. plantlets in nitrogen lacking medium at the end of the
observation day.

Without Phosphorus
In this culture medium, number of Lemna sp. plantlets stays constant for 6 days as five
and reduced to four until the end day. The number of green leaves, although seem to
increase at first, but it also reduces throughout the experiment but in a more stable
manner; and there is no apparent changes in colour of the leaves. Number of dead leaves
also in lower rate and average root length varied along the time. Since phosphorus plays
an important role in synthesizing nucleic acids, adenosine triphosphates (ATP)and
phospholipids of plasma membrane and also acts as a coenzyme in both photosynthesis
and respiration, thus it can be concluded that absence of this phosphorus resulted in poor
root formation and lower metabolism rates (leads to reduced photosynthesis and
reproduction processes).

Figure 3 : Some of the Lemna sp. plantlets in phosphorus lacking medium at the end of the
observation day.

Without Potassium
In this medium, it was noted from Table 1 that the number of Lemna sp. plantlets and
number of green leaves to increase but then dramatically fall on day 12. The number of
dead leaves ( 7 yellow leaves and 2 white leaves) also can be said to be high and the
average root length seem to be vary. Pottasium plays role in osmosis regulation but the
impact caused on growth rate is the function of potasium in opening stomata. Without
enough potassium ions to regulate the opening of stomata, the rate of photosynthesis is
affected thus directly affecting the growth of the Lemna sp. plantlets, hence explaining the
reduction of both the number of plantlets and green leaves on the end of the observation
day. Some green leaves turn into pale colour throughout the experiment indicate
symptoms of chlorosis .

Figure 4 : Some of the Lemna sp. plantlets in potassium lacking medium at the end of the
observation day.

Without magnesium
From both Table 1, it can be seen that the number of plantlets in magnesium lacking
medium decreases into three at the end of observation day. Number of green leaves seem
to decrease quiet steeply and number of dead leaves also can be said to be high, which is
about 10 leaves. The average root length seems to be varied too and the green leaves
themselves fade in colour throughout the experiment period. Magnesium is an important
macronutrient in chlorophyll synthesis and absence of this nutrient result in lower
production of chlorophyll thus reduces the rate of photosynthesis. This, without any
doubt will affect both root and shoot growth and full development of the plantlet. Some
symptoms of chlorosis can be seen on the leaves where the regions between the veins
turn yellow. And magnesium being the key to turn on other plant enzymes and being
involved in carbohydrate metabolism, absence of this nutrient slows down the growth of
plant as translocation of photosynthase is slowed down and thus affect the uptake of
other elements.

Figure 5 : Some of the Lemna sp. plantlets in magnesium lacking medium at the end of the
observation day.

Without calcium

From Table 1 it can be seen that number of plantlets stay constant for six days but fall
down to one on the final day, while number of green plant decreases across the period
dramatically. The number of dead leaves in calcium lacking medium is the highest among
all other mediums where it has 4 yellow leaves and 14 white leaves, which some of them
decompose before the end day. The average root length also deteriorates across the time
line showing that the condition of the plantlet is the worst among the bad. Calcium is the
major constituent of the middle lamella of cell walls and key for the formation of spindle
fibre during cell division. Without calcium ion, cell division cannot occur at all resulting in
the death of the plantlets in long term run. Since the permeability of the cell wall also not
regulated by calcium in this medium, so some of vital living processes fail to take place
leading towards cell dying and decaying , explaining the reduction of root length and
green leaves.

Figure 6 : Some of the Lemna sp. plantlets in calcium lacking medium at the end of the
observation day.

Without Iron
Number of plantlets in this iron lacking medium slowly increase and the same goes with
number of green leaves, but the leaves become lighter in colour across time. There is no
much variance in average root length. Absence iron affects the production of chlorophyll
leads to lower photosynthesis activity, which in turns slows down the growth of the plant.
Although iron is a micronutrient, the absence of this nutrient still give impact to plants
growth and development (slow and stunted growth).

Figure 7 : Some of the Lemna sp. plantlets in iron lacking medium at the end of the
observation day.

Without sulfur
Based on Table 1 , number of plantlets and green leaves increases without much
disruptions while only one leave was noted to turn white. Other than that, the average
length of root doesnt vary much but have its own small dramatic changes. The colour of
green leaves noted to be slightly yellowish but not fully, thus it was counted under
number of green leaves. Deficiency in sulphur halts the synthesis of certain amino acids,
nucleic acids, vitamin B and other coenzymes. This results in yellowing of leaves and
delayed maturity..

Figure 8 : Some of the Lemna sp. plantlets in sulphur lacking medium at the end of the
observation day.

Lack of all nutrients

Without any essential nutrients, the number of plantlets, number of green leaves and
average length of root goes down the slope. The vice versa happens for the number of
dead leaves and eventually none of the plantlet survive till the final day. This is because
all the metabolic activities are restricted thus the plants cant produce any food to survive
thus resulting in zero survival chance.

Figure 9 : Some of the Lemna sp. plantlets in all nutrient lacking medium at the end of the
observation day

Limitations
There are several limitations that have been identified throughout this experiment.

The Lemna sp. plantlets may be infected earlier before it is culture into the medium. This
may lead to inaccurate results as it may change the symptoms of the mineral deficiencies .

During picking up the Lemna sp. plantlet, the number of leaves was controlled to be two.
But, there are possibilities that at that very moment, the plantlet is in the middle of
having another new leaves. Thus, this will affect both the number of green leaves and
number of plantlets , leading to misinterpretation about certain mineral deficiencies.

Since water vapour formation raising difficulties in counting the number of plants, the
petri dishs lid was opened up and wiped. At this moment, the plantlets in the medium are
very vulnerable for the microorganism in the air to attack. Furthermore, this
unintentionally provides carbon dioxide for the plantlets in various amounts that may
contribute to different development.

For medium of lacking all nutrients, distilled water was used. This may alter the result as
the distilled water was placed in container and exposed to the surrounding for long time
may contain microorganisms or minerals from air.

Sources of errors
Several sources of error in this experiment were identified and steps were taken to
minimize these errors to make the result more accurate.

Floating Lemna sp. plantlets tend to clump together whenever they were given even the
slightest force. So, they were arranged to maximum distance within the petri dish to ensure
minimum competition. To avoid contamination, a pair of sterilized forceps was used gently
to place the plantlets.

The solution medium may split out when the petri dishes were brought back to the window
side. Thus, by placing cling film around the petri dish, any alteration in volume of solution
is prevented.

Conclusion
Plants need both macro (Nitrogen, Phosphorus, Potassium, Magnesium, Calcium and Sulphur)
and micro (Iron) nutrients to achieve full growth and development. Deficiency in any one of the
mineral can be spotted through the symptoms that will display on its physical appearance.
Different type of mineral deficiency results in different effects on the growth of Lemna sp.
plantlets. Thus, the hypothesis is accepted.

Further Investigation
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 are moisten to 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).

Figure 10: Set up of apparatus for further experiment (11)

References
1. Wikipedia Foundation. Last modified on 2012. Lemna. Available from
http://en.wikipedia.org/wiki/Lemna. Accessed on 3th March 2012.
2. http://www.plant-lore.com/plantofthemonth/duckweed-and-jenny-greenteeth/.
Accessed on 3th March 2012.
3. http://www.fishfarming.com/duckweed.html. Accessed on 3th March 2012.
4. Wikipedia Foundation. Last modified on 2012. Chlorosis. Available from
http://en.wikipedia.org/wiki/Chlorosis. Accessed on 3th March 2012.
5. http://www.dias.kvl.dk/plantvirology/esymptoms/symp-color.html. Accessed on 3th
March 2012.
6. http://www.evershinehydro.com/Nutrients/twotypessofnutrients.html. Accessed on 3th
March 2012.
7. http://www.victuslabs.com/6885/index.html. Accessed on 3th March 2012.
8. http://hydrophytesblog.com/?paged=4. Accessed on 3th March 2012.
9. http://newtonpost16.org/chore-macro-and-micronutrients-exporters-and-also-pricesof-macr-and-micr-nutrients/ . Accessed on 3th March 2012.
10. Gan W.Y . 2007. Biology SPM Success. Edition 4. 196-97.p.Shah Alam : Oxford Fajar
Sdn.Bhd.
11. http://www.nuffieldfoundation.org/practical-biology/investigating-effect-mineralsplant-growth. Accessed on 3th March 2012.