Botany 111.

1 Plant Morphoanatomy and Diversity Laboratory

EXERCISE 7
SECONDARY GROWTH

INTRODUCTION

Secondary growth means growth in girth or diameter. It is due to the activity of a unique tissue, the vascular
cambium, which produces new cells in the radial dimension. Secondary growth is essentially a property of
woody plants, both woody dicots and gymnosperms. Thus, an understanding of secondary growth comes
from the study of woody stem morphology and anatomy.

Secondary growth occurs as the result of lateral meristematic activity, and produces an increase in the girth
of an organ. The two most common lateral meristems in conifers and woody dicots are the bifacial vascular
cambium, that produces secondary xylem (wood) and secondary phloem, and the cork cambium (phellogen)
that either produces only cork (phellem) to the outside, or both cork and phelloderm (to the inside). In
contrast to primary meristems that have been undergoing continuous meristematic activity from the
inception of the radicle and epicotyl in the embryo, secondary meristems are redifferentiated from cells that
previously had ceased cell divisions and matured into paremchyma.

Whereas the bifacial vascular cambium is a perminant meristem, there is a succession of cork cambia
redifferentiated in the primary cortex and (later) in the secondary phloem. The secondary xylem (wood) of a
plant provides a permanent record of vascular cambial activity throughout the life of the plant. By contrast,
the secondary phloem is constantly being displaced further-and-further out into the bark, and is eventually
sloughed off with the old bark

MATERIALS

Electric or compound microscope (one per student if available)

x. s. of an old Ranunculus, buttercup root or old GYMNOSPERM Pine 1 yr old c.s (older stem)
Aristolochia root
x. s. of Muntingia calabura old stem x. s. Pine wood
DICOT Aristolochia older stem x.s Wood tangential sect. Pine
x. s. Dicot Wood WOOD radial sect. Pine
Dicot wood tangential sect.
DICOT wood radial sect.

PROCEDURE

A. The Old Dicot Root

Secondary growth occurs in many roots and usually results in the thickening of the root diameter by the
addition of vascular tissue. Initiation of secondary growth occurs when cells in the residual procambium and
parts of the pericyle begin to make periclinal divisions. Only the pericyle cells opposite the xylem points start
to make periclinal divisions. The inner layer of cells becomes the vascular cambium. The outer layer is retained
as pericycle. The vascular cambium is continuous around the primary xylem.
Exercise 7. Secondary Growth

The vascular cambium continues to divide periclinally. The daughter cells that result from these divisions
differentiate into secondary xylem cells if they divide off towards the inside of the root or secondary phloem
cells if they divide towards the outer surface of the root. After many cell divisions and cell differentiation, a
root exhibiting secondary growth might look like the one depicted in the diagram to the right.

Some roots also form an outer protective layer called the periderm which originates from the pericycle and
replaces the epidermis. The pericycle resumes its meristematic character and begins to divide periclinally
again. At this point it is called the phellogen or the cork cambium.

The cork cambium forms phellum cells (cork cells) towards the outside of the plant. These cells are dead at
maturity. They are suberized which makes the cells impermeable to water. Phellum cells in cross section
appear in neat, ordered files. The cork cambium also produces the phelloderm, a tissue consisting of cells
that are living at maturity. Not all plants have secondary growth in their roots, and many do not conform to
the developmental pattern that was described above.

Lab Work 1.

1. Examine the x. s. of an older dicot root. Observe the order of occurrence of the secondary tissues
formed by the cork cambium and the vascular cambium. Locate the (a) phellem, consisting of
suberized cork cells; (b) phellogen, a region of flattened, tangentially dividing cells; and (c) phelloderm,
a tissue inner to the phellogen, parenchymatous in nature and a few cell layers thick. The latter form
the new cortical tissue in the old root. Other than its being suberized, what other characteristics does
the phellem have? What are some of its functions?

B. The Woody Stem After Several Years of Secondary Growth

Lab Work 2.
Now let’s look at the structure of a woody stem after a few years of growth. Use an older stem of
Aristolochia cross-section for this. Find the vascular cambium. Recalling the order in which you looked at the
anatomy of young stem, locate the following to the inside of the vascular cambium:

1. XYLEM – Find the tracheary elements, parenchyma and fibers. Each of these will just be like what you
saw in the young stem. However, you will see two significant differences between young and older
stem:
a. ANNUAL RINGS: The xylem you see appears as rings of xylem. Each of these rings was
produced in a different year, and so these are called annual rings. This means, as you
well know, that the age of a woody stem can be determined by counting the number
of annual rings. Annual xylem rings appear because xylem develops differently at
different times of the growing season. The first xylem to develop is termed SPRING
WOOD. It is usually less densely packed than the SUMMER WOOD that develops later
in the growing season. Remember that xylem growth is form from the inside to the
outside, which means that the summer wood of any year will always be to the outside
of the spring wood of the same year.

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Exercise 7. Secondary Growth

b. XYLEM RAYS: Note the radiating lines running through the xylem from the center of
the stem. Xylem rays are sometimes called as wood rays. In angiosperms, xylem rays
maybe more than one cell layer thick. In gymnosperms, most rays are just one cell thick.
Rays in the vascular tissue of woody plants apparently are important for lateral
transport of substances.

2. PITH: The pith in the older stem is becoming a minor component.

Return now to the vascular cambium, and work your way to the outside, locating the following
to the outside as you go:

3. PHLOEM: As in the young stem, identify the sieve tubes, companion cells, parenchyma and phloem
fibers.
a. PHLOEM RAYS : These are the group of cells between the phloem lobes. Phloem rays
are continuous with the xylem rays. As the stem grows, cells in the phloem rays divide,
and elongate in such way that the phloem ray widens as the stem grows in
circumference (the rays are then called flaring rays). Thus, the phloem rays in older
stems have tapering form, being many cells wide toward the outside.
4. CORETX: Again, the region outside the phloem and phloem rays.
5. CORK CAMBIUM AND CORK: The outer protective layers. The cork cambium arises from the outer
layers of cortex. It continually produces cork cells, which have a higher suberin content, to the outside.
The cork cells eventually die and become part of the outer bark, or cork.
6. LENTICELS: Morphologically, lenticels are the raised areas on the surface of a mature stem that
function for exchange within the inner tissues and the outside. Anatomically, these are depressed
areas within the periderm layer that lie opposite the wide vascular rays. The concave area is filled with
loosely arranged cells called filling or complementary cells.

Return to the vascular cambium. It is a more complex tissue itself than just a bunch of dividing cells.
The vascular cambium produces two kinds of cells: long tubular cells that become xylem vessels or
tracheids inside or phloem tubes outside, and ray cells (again, xylem rays or woody rays inside or
phloem rays outside). This is accomplished by two different kinds of cells in the vascular cambium:
FUSIFORM INITIALS give rise to the tubular cells, and RAY INITIALS produce ray cells. Fusiform and
ray initials can be distinguished in a cross-section form their positions, the ray initials being continuous
with the rays, the others therefore being fusiform initials. It is not possible in a cross-section to see
the vertical dimension of the vascular cambium cells.

C. Anatomy of Gymnosperm Wood

Gymnosperms are often called ‘softwoods’. This is something of a misnomer, since some dicot trees
(‘hardwoods’) actually have softer wood than some gymnosperms.

The structure of gymnosperm wood resembles that of dicot wood, with a couple of important
differences. One difference is that gymnosperm wood has no xylem vessels, only tracheids. Another
important difference is the presence of RESIN DUCTS in xylem, phloem, and cortex of gymnosperms.

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Exercise 7. Secondary Growth

Observe a slide of a cross-section of a pine stem. By now, you should easily be able to locate the xylem
(and its parts), the vascular cambium, the phloem (and its parts), the cortex, the resin ducts, and the
cork cambium and cork.

Lab Work 3.
Look at a longitudinal section of a pine stem. This is a good place to view BORDERED PITS in the
tracheids. You have already read about the position and function of bordered pits, but be sure to look
at them in the prepared slide.

D. The Structure of Wood

As woody stems age, the oldest xylem ceases to conduct water; only the outer rings (i.e. most recently
formed) remain functional. Often a color change is associated; the center, older, non-functional region
becomes older. On the blocks of wood, you will see a dark, non-functional area called HEARTWOOD.
The outer region, still functional is SAPWOOD. In old trees, the ratio of heartwood to sapwood is high,
which means that much of the stem diameter is heartwood. The tonal contrast between heartwood
and sapwood is one of the things that adds richness and warmth to woods, and makes them highly
prized for such uses as furniture and flooring.

Look at some of the wood sections. You can easily recognize a CROSS-SECTION (sometimes called as
transverse section), because you will see entire annual rings. The other ways of cutting wood are the
RADIAL SECTION and the TANGENTIAL SECTION. Both of these are longitudinal sections, parallel with
the vascular tissue. A radial section, however, always goes through the rays (running from center out)
will exhibit their side view, and appear as line running at right angles to the up-and-down lines of the
vascular tissue and annual rings. In a tangential section, the cut is made without going through the
center. Thus, the rays are cut through, rather than along, and so we see the rays in their cross-section,
even though the vascular tissue and annual rings are still appearing in long section.

The differences between cross-, radial-, and tangential section can be easily seen microscopically. Look
at the three different sections in both a hardwood and softwood (pine). Note that the brick-like
patches are the rays in side-view, so this view must be a radial section. If, on the other hand, you see
groups of cells in a tapering or teardrop shape, then you are viewing rays from their ends, so the
section must be a tangential section. Look again at the cross-section, just to remind yourself how it
appears compared to others.

Lab Work 4.
1. Examine slides of these sections taken from an angiosperm wood. Note the characteristic feature of
each section based on the shape and the arrangement of the ray parenchyma. What are the functions
of the ray parenchyma? Sketch the three sections to show distinct features of wood.
2. Examine slides of these sections taken from a gymnosperm wood. Sketch the three sections to show
distinct features of wood.

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Exercise 7. Secondary Growth
References:

Balangcod T.D. and Buot I.E. Jr. (2011). Plant Taxonomy: A Laboratory Manual (A Revised
Edition). Philippine Society for the Study of Nature. 109 pp.
Rost T.L. (1996). Secondary Growth in Roots. Retrieved from http://www
plb.ucdavis.edu/labs/rost/Tomato/roots/secondary.html
Secondary Growth. Experimental Anatomy of Plant Development Laboratory 9. Retrieved from
https://www.ohio.edu/plantbio/staff/rothwell/pbio442-542/Lab%209%20-
%20Secondary%20Growth.pdf