OEB 52 Lab #1

Looking at Plants!

In this first lab we want to start looking at plants. How they build their bodies, how
they grow, the nature of their organs and tissues. We cant cover everything today, but
hopefully this will get us started on the road to really seeing the plant. We will focus on
topics covered so far in class. The lab needs to be hands on. Dont be afraid to pull your
plant apart to see how it is constructed and how it grows. Make sure you understand all
of the terms and key concepts listed here in the goals section before you leave the lab.
Also, be sure to illustrate these in your lab book. Drawing is a great exercise for making
sure you really are seeing the plant. Often we look at things without ever seeing things.
Becoming a botanist is a wonderful way to enrich your world; botany opens up a great
green door filled with tremendous morphological diversity and fascinating developmental
and functional features.
Goals:
1) Learn to use a hand lens, a dissecting microscope and a compound microscope.
2) Understanding primary growth: the shoot apical meristem where is it located,
what does it look like, what does it make, types of phyllotaxy, the idea of
modularity and metamers/phytomers, node/internode, axillary meristem.
3) Understanding primary growth: the root apical meristem where is it located,
what does it look like, root cap, zones of cell division and elongation, root hairs,
branching and why no phyllotaxy in roots?
4) Understanding primary growth: intercalary meristems: grass leaves, Welwitschia
leaves, as examples.
5) Understanding leaves the only plant organ we will really look at today. External
morphology: blade and petiole, simple versus compound, pinnate (and bipinnate)
versus palmate, rachis. Anatomy: trichomes, cuticle, stomata, veins, epidermis,
mesophyll (palisade and spongy parenchyma with lots of air spaces why?).
Objective 1: How to Look!
How to use a hand lens: hold the lens up to your eye, move the thing you want to look at
back and forth until it comes into focus. Task: use a hand lens to see different hairs on the
surface of a leaf.
Dissecting scope: make sure you have good illumination; you may want to vary the angle
of the illumination (if from the side), if a ring light, then it will illuminate evenly from
above. Note that you can adjust the magnification as well as the focus.

Compound scope: most important: make sure that the eyepieces are adjusted for the
spacing between your eyes! Have your TF show you how to adjust magnification and
focus. Here are some web sites with information on using compound scopes:
http://www.labessentials.com/Microscopes_Compound_Basics.htm
Here is a youtube on how not to use a microscope: http://www.youtube.com/watch?
v=vRIA04U796E&feature=related
And heres a serious one with more info:
http://www.youtube.com/watch?v=L6d3zD2LtSI&feature=related

Objective 2: Shoot apical meristems

First Grab a plant and locate the shoot apical meristem(s). How many are there? What
does the shoot apical meristem do? Try to dissect one to find the smooth dome of tissue
that is the real meristem (persistent population of embryonic cells).
There are three basic configurations for leaf primordia (and thus mature leaves) in
relation to the shoot apical meristem: opposite (i.e., produced in pairs), whorled (i.e.,
produced three or more at a time (= three or more at the same node), and alternate (i.e.,

produced in a spiral). Can you convince yourself that an alternate arrangement of leaves
on a stem is the product of leaf primordia being produced in a spiral around the shoot
apical meristem?
For opposite phyllotaxy: successive pairs of leaves can be rotated by 90 degrees,
producing a pattern we refer to as decussate, or all pairs of leaves can be produced in the
same plane. In terms of intercepting sunlight, which would be better?
We can use several methods to characterize spiral phyllotaxy. One way is by the number
of leaves that are produced before the spiral gets back to the same starting point and by
the number of times that you have to go around the stem before you go back to the same
starting point (i.e, until you have a leaf that is directly above or below the one you started
counting from).
Have your TF show you movies of living meristems. What does P0 mean? See if you can
recognize the spiral pattern of primordium initiation.
Now lets think about the concept of modularity and the idea of a metamer (also called a
phytomer) which is a repeating morphological unit produced by the shoot apical
meristem. In most angiosperms: the shoot metamer is a node (point where the leaf is
attached) and the internode (elongated stem between nodes), plus an axillary meristem
produced in the axil between the base of the leaf and the stem. See if you can identify
all of these parts that make up the angiosperm metamer on at least three plants in the lab.
How is the internode produced? Why does the plant make axillary meristems? Along a
stem that no longer has leaves (i.e., these have already fallen off), where would you look
for axillary meristems relative to the scars left by the fallen leaves?
Objective 3: Root apical meristems
Dig up some roots, can you find the root apical meristem? How does it differ from the
shoot apical meristem? Can you find root hairs? Why are they not produced right next to
the root apical meristem? What does the presence of root hairs tell you about where
elongation is (and is not) occurring?
Where are root branches formed? Why are they not produced by the root apical
meristem? Why do roots lack phyllotaxy? How does branching differ between roots and
shoots?
Objective 4: Intercallary meristems
To complete our survey of meristems responsible for primary growth we need to cover
intercalary meristems. These are meristematic regions that occur in the middle of mature
tissues i.e., regions of cell division and expansion that push forward mature tissues. For
example, the leaves of Welwitschia are formed by an intercalary meristem at the base of
each leaf that produces new tissues that propels forward the older leaf tissues (of these

strap shaped leaves); grass leaves are similar. This results in the youngest tissues being at
the base and the oldest tissues at the tip.
Objective 5: Leaves
Finally, lets look at leaves. First lets name the parts of a leaf and the two basic types:
simple leaves have a single blade or lamina (the flat photosynthetic part) and a rounded
or flattened petiole which connects them to the stem; compound leaves have their
photosynthetic area divided up into multiple leaflets. Compound leaves can be pinnate or
palmate, and they can be once-pinnate or twice-pinnate. We give a special name to the
structure to which the leaflets are attached: the rachis. It is like a petiole in terms of its
function, but we give it a special name. Question: how do you distinguish a compound
leaf from a stem segment with leaves? (hint: what is NOT found in the axils of the
leaflets???)
For fun: why simple versus compound leaves? Think of advantages and disadvantages
associated with each type?
Now lets look at the surfaces of leaves: try to find leaves with different types of hairs
(trichomes) and waxes. Think about why leaves might want to be reflective? Where are
the stomata located? Be sure to look at the stomata on more than one type of leaf. Do
they differ in size? Number? Location?
Then make a cross section of a leaf (or look at a prepared slide, but please first try to
make your own, its much more fun) what do you see? Is the leaf solid or does it have
air spaces? Where are the chloroplasts (green cells!) located? Can you find chloroplasts
in the epidermis? (the answer depends on which type of leaf you look at, yes in ferns, no
in angiosperms, for example). Do stomata have chloroplasts? Turns out yes, in all cases.
We will talk about why when we discuss how stomata work. Why do leaves have air
spaces? Notice that these photosynthetic parenchyma cells are arranged differently near
the upper surface of the leaf (palisade parenchyma) versus the lower surface (spongy
parenchyma). Can you see the cuticle? It is a layer on the outer surface of the leaf that
helps prevent water loss. For now just look at the veins and think about what roles they
place (mechanical support, delivery of water and nutrients, export of the products of
photosynthesis). We will talk about the vascular tissues soon in class.
General information useful for making and staining plant material:
Staining Reagents
Toluidine Blue O: General metachromatic staining.
Hematoxylin: Nuclear staining.
KI: Starch staining.
Trypan Blue: Dead cell staining.
Cotton Blue: To stain the chitin in fungis cell wall.
These stains can be removed by the Glycerol: water = 1:1 solution