EXERCISE 1: CONTROL OF SENESCENCE

ABSTRACT

Growth is a very crucial biological process in all organisms, as well in plants, which we
universally consider as primary producers. Plant growth is

INTRODUCTION

Growth is a very crucial biological process in all organisms, as well in plants, which we
universally consider as primary producers. Plant growth is generally described as the increase of plant
volume and/or mass, which may or may not be associated with the formation of the new structures and
organs, and involves cell division, expansion, and differentiation (Brukhin and Morozova, 2011). In an
article by Brukhin and Morozova (2011), they mentioned that plant growth is mainly attributed to the
increase of cell numbers and size.

Multicellular organisms such as plants have regions or loci that are called meristem (Brukhin and
Morozova, 2011). Meristems are embryonic tissues considered as center of cell division and growth and
stays throughout plants’ life (Morrow et al, 2021). There are three main kinds of meristems: Apical
meristem, which occurs at growing shoot and root tips, designated as Shoot Apical Meristem (SAM) and
Root Apical Meristem (RAM) respectively; Lateral meristem, which is responsible for producing the cells
that makes up plant stem tissues; and Intercalary Meristem, which only exist in monocots, occurs at the
stem nodes (Gilbert, 2000).

There are different ways to determine the growth of a plant and its organs, such as measuring
height, width, mass, cell number, and content level of protein or other essential substances (Brukhin and
Morozova, 2011). However, the growth rate of a plant and its organs vary widely, and plant growth is
rarely indeterminate (Grime and Hunt, 1975; Yin et al, 2003). However, growth duration of most
organisms, especially plants, can be divided into three phases, namely, lag or early growth phase,
exponential or rapid growth phase, and declination or saturation or stationary phase, and therefore
plant growth typically exhibit a “S” or sigmoid curve (Lieth et al, 1995; Yin et al, 2003, Fig .1)

Figure 1. Illustration of a sigmoid biological curve. Lifted from Sun et al (2015)

 This exercise aimed to:

1. Determine the locus of growth in the root and leaf;

2. Analyze the different phases of growth and the growth rates of chosen plant
species; and
3. Estimate the relative growth rate of a given species.

METHODOLOGY

Locus of growth in the root

Mungbean (Vigna radiata) seedlings were soaked in water and allowed to germinate for 48
hours or until the radicle grew up to 2 cm. Five (5) best seedlings were selected, blot dried their roots
then made 5 marks on them at 1mm distance from the tip, and then let the ink dry. The seedlings were
then placed on a moist paper, rolled the paper then placed inside a plastic bottle with a small of amount
of water. The bottle was placed inside a cabinet or any dark area and allowed the seedlings to grow for
24 hrs. After 24 hours, the distance between the marks were measured and recorded.

Five (5) marks on The seedlings were The bottle was

Mungbean the roots at 1 mm. placed on a moist placed inside a Distance between
Five seedlings were
seedlings were paper, rolled the cabinet or any dark the marks were
selected and blot- Z -5
soaked for 48 paper, and placed area and allowed measured and
dried their roots. Z-4
hours. Z-3 inside a plastic the seedlings to recorded.
Z -2
Z -1 bottle. grow for 24 hours.

Figure 1.

Locus of growth in the leaf

Three (3) replicates of young leaves were chosen and then labeled. Clear plastic sheet was laid
on top of the leaves and traced their margin using a marker. The leaf outlines were then laid on a graph
paper and traced the outline of the grids. Three (3) regions were made in the outline leaf and counted
the squares in each region and then recorded the data.

Leaf margin of the leaves was

measured by laying a clear
plastic sheet on top. Three regions were made in
The leaf outlines were then
Three young plant leaves were the outline leaf and counted
laid on a graph paper and
chosen and labeled. the squares in each region and
traced the outline of the grids.
then recorded the data.

Figure 2.
Growth of an intact plant

Seeds of the chosen plant were planted and germinated. At least six (6) good seedlings were
chosen and grown under normal conditions until maturity. The height of the plants was measured and
recorded daily.

Seeds were planted and germinated.

Chosen seedlings were grown under normal conditions until maturity.

The plants’ height was measured daily and recorded.

Figure 3.

RESULTS AND DISCUSSION

Table 1. Length of each zone in mungbean root tip after 24 hours of incubation in the dark
Length (mm)
Zone Number Mean
Rep 1 Rep 2 Rep 3 Rep 4 Rep 5
1 10 14 5 8 4 8.2
2 1.2 1 1 1.1 1 1.06
3 1.1 1 1 1.1 1.1 1.06
4 1 1 1 1 1 1
5 1 1 1 1 1 1
total 14.3 18 9 12.2 8.1 12.32

20
A
18 B
16
14
12 R3

10
8
6
R5
4
2
0 R1
Zone 1 Zone 2 Zone 3 Zone 4 Zone 5 R2
Replicate 1 Replicate 2 R4
Replicate 3 Replicate 4
Replicate 5
Figure 1. (A) Line graph of the length of each zone in mungbean root tip after 24 hours of incubation (B) Mungbean seedlings
after 24 hours of incubation
 In this first part of the exercise, we observed the change of length of different zones of the root
of Vigna radiata seedling after 24 hrs. As shown in Table 1. The zone of that the most obvious
elongation is the zone 1 or the tip most part. Zone 2 and 3 has the most minimal elongation and the rest
of the zones remained unchanged.

As mentioned earlier, multicellular organisms such as plants, has distinct zones where cells
actively proliferate, which are called meristems, and the meristem that produces cell in root tips for
continues root growth is called Root Apical Meristem (RAM), a kind of apical meristem tissue (Brukhin
and Morozva, 2011). Generally, RAM is composed of five major tissues, namely epidermis, ground
tissue, which consists of cortex and endodermis, stele, which consists of pericycle and vascular tissue,
root cap, and quiescent center, which are less active dividing cells but important in maintaining the
surrounding cells (Terpstra and Heidstra, 2009). On a lateral view, root cells can be divided into three
distinct developmental zones, namely, meristematic zone (MZ), elongation zone (EZ), and differentiation
zone (DZ) or maturation zone in some literatures (Petricka et al, 2012). The meristematic zone is an area
of active cell division takes place to generate a pool of cells that will later on elongate and differentiate.
Meristematic zone also consists of Stem Cell Niche (SCN), Proximal Meristem (PM), and Transition Zone
(TZ) (Fig 2) (Jose and Li, 2013) in elongation zone, cells tend to divide less and focus more on increasing
their size through elongation while in differentiation zone, cell elongation becomes scarce and cells
differentiate into cells with specific functions such as root hairs (Petricka et al, 2012; Jose and Li, 2013).
 Figure . Labeled tissues of Root Apical Meristem (RAM). Major developmental zones along the longitudinal axis of a root are
marked in color bars. (EZ) elongation zone; maturation zone, also known as Differentiation Zone (DZ); (MZ) meristematic zone,
which consists of: (SCN) stem cell niche; (PM) proximal meristem; TZ, transition zone. Lifted from Jose and Li (2013)

Based on the results, since Zone 1 has the most apparent change in length, we could consider
that Zone 1 contains the meristematic zone and elongation. Zone 2 and Zone 3 did have a little change,
so they could still be part of the elongation zone. But for Zone 4 and 5, since there there’s no change in
length and root hairs already emerged in these two areas (Fig. 3), they are part of the differentiation or
maturation zone.

Table 2. Final area of each region on the Ocimum basilicum young leaf
Initial Leaf Area (cm2) Average Final Leaf Area (cm2) Average
Zone Replicate Replicate Replicate area Replicate Replicate Replicate area
1 2 3 (cm2) 1 2 3 (cm2)
tip 0.75 0.75 0.75 0.75 0.75 0.75 1 0.83
mid 1.75 1.5 1.75 1.67 1.75 1.75 1.75 1.75
base 1 1 1.25 1.08 1.25 1.25 1.75 1.42

Diagram

In this second part of the exercise, we observed the change of leaf area of a young leaf after 5
days. As shown in Table 2, there is very minimal overall change of average area but among the zones,
the base has the highest change of average area. The base zone increases in area of 0.34 cm2 while both
tip and mid zones increased in area of 0.08 cm2.

Unlike SAM and RAM that exhibits indeterminate growth, the growth the leaves exhibit is
determinate (Ichihashi and Tsukaya,2015). Cells that develop and grows into leaves are produced by leaf
primordia, which can be found in SAM region (Larren, 2017). But the proliferation of the cells in leaf
primordia is restricted and spatially separated from SAM (Ichihashi and Tsukaya, 2015). Esau (1977)
stated in her book that organogenesis in leaf primordium depends on many meristematic tissues
including plate meristem and marginal meristem. Plate meristem produces cells that divide in anticlinal
direction which contributes to the leaf growth while marginal meristem, which is located at the edge of
the leaf and separates the adaxial and abaxial side, produces cells that comprises the tissue layers within
the leaf (Esau, 1977). These leaf meristems dictate the orientation of the cell division plate then results
to different compartments, namely leaf blade and petiole, and determine the gradient in developing
leaves where the cells switch from proliferation to differentiation (Donnelly et al., 1999; Ichihashi et al.,
2011). This gradient point is said to remain at a constant position and diminishes abruptly upon leaf
maturation (Kazama et al., 2010; Andriankaja et al., 2012). This also means that at a certain period, the
leaf meristematic regions cease cell proliferation and will be localized somewhere in the leaf base and
petiole (Ichihashi et al., 2011). This tells us that the production of new cells in leaves occurs in the base
and the cells in tips are the older cells. This coincides with the results of the exercise, as the base zone
has the increased the most among all the zones in terms of estimated area.
Figure _ (A) Developmental stages of Arabidopsis thaliana leaf showing the proliferative region (red). (B)
Left image: A leaf primordium at 7 days after sowing with cell lineages indicated by blue staining. The
middle and right images indicate the spatial differentiation of leaf meristems. Lifted from Ichihashi and
Tsukaya (2015)

Table 3. Periodic height of soil grown Zea mays plants

Plant Age Measurement (cm)
Mean k
(Days after emergence) 1 2 3 4 5 6 7 8 9 10
1 1.9 3.1 4 4.8 5 4.9 3.8 1 2.5 4.5 3.55 -
7 17 17 16 15.5 21.5 15 16 15 12 13.5 15.85 2.58
14 25.5 22 21 22.5 25 22 23 24 20 22 22.7 3.03
21 30 30 24 23 35 25 26 27 26 23 26.9 3.23
28 42 43 30 32 51 36 39 40 34 32 37.9 3.59
35 48 48 40 36 56 46 44 46 45 41 45 3.77
42 84 77 53 49 84 65 73 65 65 59 67.4 4.18
46 95 85 56 52 87 70 76 71 70 64 72.6 4.26
 H vs t lnH vs t
100 5
90 4.5
4
80
3.5
70
3
60 2.5
50 2
40 1.5
30 1
0.5
20
0
10 Day 1 Day 7 Day Day Day Day Day Day
14 21 28 35 42 46
0
Day 1 Day 7 Day 14 Day 21 Day 28 Day 35 Day 42 Day 46
Rep 1 Rep 2 Rep 3
Rep 1 Rep 2 Rep 3 Rep 4 Rep 4 Rep 5 Rep 6
Rep 5 Rep 6 Rep 7 Rep 8 Rep 7 Rep 8 Rep 9
Rep 9 Rep 10 Rep 10
Figure 5 (A) Line graph of the periodic height of Zea mays plants against number of days (B) Line graph of the antilog
transformed periodic height of Zea mays plants against number of days

In this last part of the exercise, the growth of intact plants is observed and determined if it
exhibits the sigmoid type of growth curve. As shown in Fig._A, which is the plotted graph of the length of
ten corn replicates against the number of days, the resulting curve looks somewhat of an S-curve or it
somewhat resembles the sigmoid curve.

Assuming that it generally exhibits an S curve, we concur that from Day 1 to Day 21 the corn
plants is in lag phase or the initial phase of corn growth. The growth stage of the corns in this phase
ranges from V1 to V10 where the corn plants are focused on height growth, emerging of leaves, and
growth of nodal roots (Ciampiyti et al, 2011; Nleya et al, 2016). The exponential or log phase or rapid
growth rate of the corn plants starts from Day 21 to Day 42. The growth stage of the corns in this phase
ranges from V11 to V14+ where the corn plant will undergo rapid growth and fast elongation of stem
(Ciampiyti et al, 2011; Nleya et al, 2016). Lastly, the stationary or the declination phase starts from Day
42 until the last day of recording which is Day 46. The growth stage of the corns is mostly likely the VT or
the tasseling stage, where the corn plants are already in or almost reaching their full height and will now
start to transition into reproductive stages (Ciampiyti et al, 2011; Nleya et al, 2016). One sign that a corn
plant is already in a tasseling stage is the presence of tassel, which can be observed in one of the
replicates and is shown in Fig _ below. In Figure _B, it shows the plotted graph of antilog transformed
corn height against the number of days. Since the equation used for the computation is a linear
equation, a straight-line graph is expected. From Day 1 to Day 7, it seems to be showing an exponential
growth but from Day 7 onwards, the graph seems to resemble a straight line so it can be said that the
overall trend of the graph is linear. With this data presented, we can conclude that the growth of corn
plants does exhibit the sigmoid growth curve.
 Figure _ The presence of tassel in one of the corn plants

Figure _ Different growth stages of Zea mays plants

CONSLUSION

The growth of a plant can be observed by measuring parameters of the whole plants or some
specific plant organs. We can also observe the growth of different loci of plant organs. We observed the
growth of Vigna radiata seedling root and determined that its locus of growth is the tip most part of the
root or as we designated in this exercise, the Zone 1. We also observed the growth of the leaf area using
a young leaf of Ocimum basilicum and it was determined that it’s locus of growth is the base zone.
Lastly, this exercise also determined that Zea mays does exhibit a sigmoid type of growth curve.

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