The Effect of Nutrient Deficiency on Tomato Plant Development

By: Darian C. Thomas

ABSTRACT
The purpose of this study was determine if a plant treated without nitrogen, phosphorus,
or with distilled water would have an effect on the biomass and standard chlorophyll content of
the plant. Tomato plant roots were exposed to a water basin in a hydroponic retriculator which

exposed the roots to solution that contained either no nitrogen, no phosphorus, or distilled water,
depending on their treatment. The plants grew for four weeks under artificial lighting in a lab.
Plants that lacked Nitrogen and distilled water displayed extremely stunted growth when
compared to a control treatment which was supplied with complete nutrients. All treated plants
displayed a lower biomass than the control group. The standard chlorophyll contents of the
treated groups did not vary greatly from the control. Such findings imply that a lack of
macronutrients in a plant can have adverse effects of its growth and ability to photosynthesize
efficiently.

INTRODUCTION
In this experiment plants were deprived of the nutrients nitrogen, and phosphorus and
grown in lab over a series of 4 weeks. Our purpose was determine if such deficiencies would
result in changes in the plants growth and development. In plant development, nutrients are
acquired through the soil to travel through the roots into the phloem so that the stems and leaves
have sufficient nutrients(Kosinski,2015). Nitrogen and phosphorous are mobile primary
macronutrients in plants that play a vital role in growth and regulating hormones. These nutrients
are mobile because when the plant originally detects the deficiency, it can easily draw nutrients

from its older parts to accommodate the loss. In plants, nitrogen is fixed by bacteria, transformed
into ammonium or nitrate, and transported via the phloem to the plants stem and leaves.
Nitrogen then allows rapid growth, seed production, and stores itself in chlorophyll for
photosynthesis to occur (Prince, 1999). Additionally, this nutrient is responsible for up to 5% of
a plants mass. Nitrogen deficiency is typically assumed when foliage is yellow in appearance
and the plants growth is stunted (Salisbury and Ross,1992,pg.130)). On the molecular level, a
lack of nitrogen causes a lower presence of cytokinins, and an increased concentration of
abscisic acid, which is responsible for leaf development, as well as stem elongation (Bergmann,
1992,pg.88). Phosphorus is normally present in plants as phosphate, and it is effective in
blooming, growth rate, and oil formation. Plants that lack phosphorus can display purple veins,
stems, and foliage, along with thin stems (Bergmann,1992,pg.98). In general,nutrient deficient
plants display stunted growth and yellow ,wilted foliage known as chlorosis.
In this lab, biomass and standard chlorophyll content (SCC) were examined as the
dependent variables. Biomass is simply the entire mass of each plant and SCC is the amount of
green pigment present in leaf chlorophyll. Several tomato plants were planted so that one group
lacked nitrogen, another would lack phosphorus, and another contained no nutrients as it was
treated with distilled water. Based on the fact that nitrogen is responsible for rapid growth, and
involved with chlorophyll development, we expected that the nitrogen deficient plants would
exhibit stunted growth, and lighter leaves when compared to the control group. Additionally, the
typical responsibilities of phosphorus in blooming, plant maturation, and oil formation led us to
believe plants with this deficiency will show wilted leaves, chlorosis, as well as a purple, wilted
stem. Distilled water lacks the normal nutrient content of tap water due to its pure qualities,
therefore, we expected that the plants watered with distilled water would exhibit extreme stunted
growth and wilted foliage. Stunted growth in each treatment would mean a lower biomass when
compared to the standard control group. Yellow foliage, or chlorosis would be confirmed by
lower SCC values.After forming such predictions, we completed the experiment to examine if no
change in biomass or standard chlorophyll content would be observed in the nitrogen deficient,
phosphorus deficient, and distilled water treatment groups.

MATERIALS AND METHODS

We prepared the plants for the study by randomly selecting plants from a sample of
tomato plants. As directed by our out of manual procedure, we removed the soil by shaking it off
into the trash and subsequently washing the roots using tap water. The roots were then carefully
patted dry. The plants were then separated with some being replanted, and others being examined
for their standard chlorophyll content. The plants were planted in slotted, black cups that were
scrubbed and washed. When planted, the roots were threaded into the slots with an inoculating
loop. Next, clay pellets were added to the cup to allow the stem to remain upright. The plants
were suspended over hydroponic retriculators with the roots falling into the underlying fluids in
the retriculators. Over a four week time span, the roots were exposed to the different deficiencies

via varying solutions flowing over the roots in the retriculators. The tomato plants that were not
replanted were weighed to determine their biomasses on a scale. 5 g of each plants were
removed from their bases and crushed along with 15 mL of acetone using a mortar and pestle.
The leaves were crushed during 50 rotations, and the green product was poured into a 50 mL
beaker. Next, we filled a centrifuge tube of the way with the solution in the beaker. The
mixture was centrifuged for 5 minutes via 2000 rotations per minute. Once the tube was
successfully centrifuged, we measured its absorbency using a spectrophotometer set to a
wavelength of 663 nm. To carry out this procedure a cuvette was filled with pure acetone and
placed in the spectrophotometer to blank the apparatus. Then, a cuvette with the centrifuged was
added and the absorbance was recorded. This value was used to calculate SCC.
After a month, the plants were harvested from the retriculators and observed for any
qualitative variances between the control group and the treatments. The clay pellets were poured
into a bin and the roots were rinsed and patted dry using the same procedure as before they were
planted. The entire plant was then weighed in a weigh boat on a scale. Because some plants
exhibited extreme growth, the plant was cut into many pieces and then weighed so that it would
fit into the weigh boat. Next, 5 g of the leaf blades were carefully selected so that they
represented the plant and crushed with 30 mL of acetone in a mortar and pestle. This samples
SCC was calculated using the same procedure. Because the initial absorbance displayed a value
higher than one, the centrifuged extract had to diluted to get an accurate reading. To dilute the
extract, 0.5 mL of extract was removed from the top of the centrifuge tube and added to a 10 mL
graduated cylinder along with 9.5 mL of acetone. This diluted solution was transferred back and
forth between a cuvette before being added to the spectrophotometer to observe absorbency. The
SCC and biomass values were recorded and gathered for data observation.

RESULTS
Table 1. Biomass Comparison for Complete versus -N, -P, Distilled water treatments
Treatment

Distilled

-Nitrogen

-Phosphorus

Complete

Mean (g)

20.38

32.32

42.45

70.73

Chi Squared v.
Complete Trtmt

16.710

15.629

7.822

P-value
(trmt. vs
complete)

4.36x10-5

7.71x10-5

0.005

Each treatment was compared to the complete control group. The data was unpaired. The
distilled treatment group displayed the lowest biomass amongst all of the plants. Phosphorus
plants displayed the highest value for biomass when compared to the control Complete group.
Table 2. SCC Comparison for Complete versus -N, -P, Distilled water treatments
Treatment

Distilled

-Nitrogen

-Phosphorus

Complete

Mean (mg/g)

0.329

0.364

0.195

0.735

Chi Square v.
Complete

1.243

6.329

9.900

P-value (trtmt. v.
complete)

0.265

0.012

0.0017

Of the treatment group, -Phosphorus displayed the lowest value for SCC while the -Nitrogen
group displayed the highest value. The -Nitrogen and Distilled water treatments were greater
than the -Phosphorus group, however, they deviated greatly from that of the control group.
Table 3. Initial values for Biomass and SCC
Treatment

Distilled

-Nitrogen

-Phosphorus

Complete

SCC (mg/g)

0.211

0.19

0.22

0.244

Biomass (g)

15.03

16.83

10.13

13.81

Plants for the initial value were randomly selected from a given amount of tomato plants.
DISCUSSION
In this study, we found that the biomasses for tomato plants treated with -Nitrogen,
-Phosphorus, and -Distilled water were significantly less than the biomass of the control. Of this
data, the -Nitrogen and distilled water treatments showed a small deviation from each other. This
is most likely due to the tendency of plants treated with distilled water to behave as if it were
experiencing an extreme nitrogen deficiency. The trend of lower biomass can be explained by a
lower rate of cell production and stunted growth. Because, nitrogen and phosphorus are both
macronutrients, it is hard for a plant to maintain its optimum nutrient capacity and optimize
water flow to prevent cell death when they are absent. Our prediction that there would be a
change in biomass calculation before and after 4 weeks of growing in lab in all treatments when
compared to the control was confirmed. We were able to reject 3 null hypotheses in the biomass
experiment as the p-values in all treatments displayed a value less than 5% (Table 1). The
hypotheses were that for the -Nitrogen, -Phosphorus, and Distilled water treatment groups, there
would be no deviation between the mass of the complete group and each respective group.

Furthermore, the -Phosphorus treatment showed the highest biomass result of the deficient
treatments. This is most likely attributed to the fact that in tomato plants lacking phosphorus, the
plant can still grow a great bit, however, it will be thinner and have fewer leaves than a healthy
plant (Bergmann, 1992, p.98). When this data is compared with their initial averages for
biomass, we see that all plants grew. Nevertheless, the -Nitrogen and Distilled water groups only
vary slightly from their initial values for biomass. This was expected given that nitrogen is
imperative for the synthesis of cytokinins, which encourage cell division.
In the standard chlorophyll content experiment, there were a few unexpected results.
Chlorophyll is responsible for the green pigment in foliage. A typical display for plants that lack
phosphorus is dark green leaves. Therefore, we would expect that phosphorus would have a
closer SCC to that of the control group than the others, however, it deviated the most (Table 2).
In fact, in Table 3 we see that the initial SCC value for phosphorus was 0.22. This implies that
this treatment group so little to no change in content. This is possibly due to the fact that the
plants used in this experiment had a sufficient amount of this nutrient prior to the beginning of
experimentation, and was able to recycle phosphorus during the four week time frame. This may
explain why there was such a small deviation for the control group in biomass as well.
Nevertheless, we were able to reject the null hypothesis that there would be no variation between
the -Phosphorus treatment SCC and that of the control. The -Nitrogen and Distilled water
treatment saw an increase in SCC when compared to the values in Table 3, however, they were
significantly lower than the SCC of the control. We were able to reject the null hypothesis stating
that plants lacking nitrogen would have a SCC that was the same of the Control group as the pvalue was less than 5%. However, we failed to reject the null hypothesis for the distilled water
treatment due to the fact that some ending data was not able to be obtained due to plant death.
Visually, it took two weeks of the plant growing in the retriculators for a noticeable
variance to be seen in their qualitative development. Within three weeks, the distilled water
treated plants were the shortest of all treatment and displayed more signs of chlorosis than any of
the other treatments. Also, the control saw an extreme amount of growth, so much that they
outgrew the framing and began to fall over the edge of the retriculators. Because of the thin
stems in the phosphorus deficient treatment, we saw that the plants leaned in different directions
to find support for the railing that contained them. Nitrogen deficient plants appeared to have
lighter leaves than that of the control group, and displayed a height between that of the
-Phosphorus and distilled treatments.
As stated in the results section, the plants that were randomly selected to be used in this
experiment were fairly healthy. This means that in terms of initial nutrient values, they may have
had enough nitrogen and phosphorus to sustain them for four weeks in the potted soil.
Furthermore, some soil may not have been completely rinsed from the roots, therefore, roots may
have still drawn nutrients from the minimal amount of leftover soil. Also, in the lab the separate
treatments were grown in separate retriculators, under separate lighting. This may have skewed
the results in the manner that some treatments may have experienced a different intensity of light
than other cohorts. This would have the greatest effect on the results as light is important for the

metabolic processes that occur in leaves. Additionally, planting and harvesting was carried out by
several students on different days, therefore, variances in technique and time of harvest may have
had an adverse effect on results.
Finally, an unpaired median data test was used for data calculation in this experiement as
there was a huge data pool and different plants grow at different rates. Also, because many plants
were examined by several freshman Biology lab sections, it was nearly impossible to keep up
with which group tested which plants. Also, the plants used to obtain initial data for biomass and
SCC were not the same as the plants that were planted in the retriculators. Therefore, different
individuals were studied before and after experimentation. Future replications of this experiment
can be enhanced by repeated the same treatments in potted soil plants, as in general, plants are
grown in soil. Also, artificial lighting may be responsible for less dramatic variances in results.
To avoid this error the plants should be placed in direct sunlight over the time frame. Future
investigations might also entail an in depth look at the effect the mobile and immobile nutrient
deficiencies will have on plant development. This could be accomplished by pairing -Nitrogen
and -Phosphorus treatments together with one on the immobile nutrients.

LITERATURE CITED
Bennett, W. F. 1994. Plant nutrient utilization and diagnostic plant symptoms. Pp. 1-7 in W. F.
Bennett (Ed.), Nutrient Deficiencies and Toxicities in Crop Plants. APS Press, St. Paul, MN.
Bergmann, W. 1992. Nutritional Disorders of Plants: Development, Visual and Analytical
Diagnosis. Gustav Fischer Verlag, New York.
Kosinski R. Plant Water and Nutrient Relations. Biology 1100 class handout. Clemson
University.
Roorda van Eysinga, J. P. and K. W. Smilde. 1981. Nutritional Disorders in Glasshouse
Tomatoes, Cucumbers and Lettuce. Centre for Agricultural Publishing and Documentation,
Wageningen, the Netherlands.
Prince, Robert. "Plant Nutrients." Plant Nutrients. North Carolina Department of Agriculture, 09
July 1999. Web. 21 Apr. 2015. <http://www.ncagr.gov/cyber/kidswrld/plant/nutrient.htm>.
Salisbury, F. B. and C. W. Ross. 1992. Plant Physiology, 4th Ed. Wadsworth Pub. Co., Belmont,
CA.

Wilcox, G. E. 1994. Tomato. Pp. 137-141 in Bennett, W. F. Nutrient Deficiencies and Toxicities
in Crop Plants. APS Press, St. Paul, MN.