Just a reminderplease be respectful of your fellow

students....

If you are chatting too much then students around you will
not be able to hear me....

If you already know something I am talking about does

not mean that someone nearby knows it.......

Please also refrain from texting and using your phones

(computers)........

Lots of new material coming that require your attention!

Thanks!
This lecture material will not be on the midterm next Tuesday Feb 21!

Dont forget midterm next Tuesday Feb 21 !

All you need to bring is a pencil or pen for the exam.

2 review sessions Mon Feb 20, 5-6 pm and 6-7 pm, 338 Koshland

Please come to the front door of Koshland Hall (immediately North of

here) and GSI will let you in as doors are locked. If you are not there by
5:05 or 6:05 there will be no one to let you in, so be on time..

We do not expect many of you will need to come to reviews J

If you need extra help go to GSI office hours !

Bring your questions to Discussion sections too J

 Plant hormones

Hormones are chemical signaling molecules

Plants vary in size from a few centimeters to many meters in

height, and all plants operate with the same principles.

How do different parts of the plant communicate with each

other?
Plants do not have moveable appendages (arms or legs etc).
Plants do not have a nervous system.

However, plants do have a vascular system.

Plants use a slower (versus direct movement or signaling via a

nervous system) mechanism of communication via moving
small chemical messengers, called hormones, through the
vascular system and cell to cell.
 Hormones
Hormones are chemical signals that are produced in one part of an organism
and function in another. Hormones produced at the top of the plant affect the
pattern of growth at the bottom of the plant and vice versa !
Discovery of a plant hormone is attributed to Darwin who first observed that
plants bend towards light
Charles Darwin discovered that there was something at the tip of the plant that was
responding to light because if you cover the tip with foil there was no response.
However, if you cover the rest of the plant with sand and just leave the tip exposed,
there was still a bending towards the light!

TINFOIL CAP

LIGHT LIGHT

FINE SAND

SOIL
One biological system that Darwin used was the coleoptiles of grasses.

A coleoptile is the pointed protective sheath covering the emerging

new leaf in monocotyledons.

Darwins experiments suggested there was something at the tip..

Darwin (and his son Francis) were the first
to study the bending of plants towards
light. They also suggested there was an
influence at the tip that caused the
bending lower down.
Fritz Went proved there was a growth promoting
substance produced at the tip..

Fritz cut off the tip of the coleoptile and placed in on

an agar block to allow substances to diffuse from the
coleoptile into the agar. (Here the substances are
drawn as pink)

He then put the agar block on the top of a cut-off

coleoptile either

a) Symmetrically, in which case the

coleoptile grew upwards.

b) Asymmetricaly, in which case the

coleoptile grew in a curved direction.

Thus, there must have been something in the original

tip that went into the agar to cause bending and
growth in response to light.

He called the substance auxin (growth in Greek).

 How does bending occur in response to light?

The plant hormone auxin causes cells to take up water and thus expand.

Light stimulates auxin production on the opposite side away from the light source,
so there is more auxin on the darker side of the coleoptile.Thus, there is more
expansion (due to water uptake) of the cells on one side versus the other side.

Thus, bending is due to asymmetric cell expansion in response to auxin and light.
 Bioassay = the testing of a biological system for a response to a chemical substance.

At low amounts (concentration) of a substance there is less response than at high

concentration.

Response

Concentration
Imagine the response is the amount of bending and the concentration is the amount of auxin.
Scientists used the bioassay of coleoptile bending to chemically
identify auxin. At low auxin concentrations the angle of curvature of
the coleoptile was less than at high concentrations of potential auxin
compounds in the agar block.

Attendance poll:

Can you guess where auxin was found in large quantities to allow its
first identification?

A. Plant juice
B. Animal stomachs
C. Animal urine
Auxin was found in highest concentration in animal urine (!) and was
then purified to allow its chemical structure to be known.

How would you know that urine had a lot of auxin?

You could do a bioassay to compare the amount of coleoptile

bending that occurred with plant juice, versus stomach juice,
versus urine J
Auxin is made at the top of a plant.

Does auxin just stay at the tip and influence growth below?

What is the direction of auxin flow?

Auxin flows downward from the tip of the coleoptile through

the stem to the roots.

How did they figure this out?

Auxin from a cut segment of the coleoptile will flow into an agar block
in one orientation, from the apical (A) end on top to the basal (B) end on bottom

The agar block resulting from placing the coleoptile segment in the A to B direction
will cause growth or bending of the coleoptile because the auxin flowed into the agar.
(The reverse experiment, placing the cut segment in the reverse orientation,
B to A direction, will not yield auxin in the agar block.)
Auxin flows from top to bottom, so only in orientation 1 will the auxin be able to
flow out and into the agar block. Auxin is colored pink below.
An important consequence of polar (top to bottom) transport of auxin is called
apical dominance.

The apex is dominant and keeps buds that are lower down from sprouting.

{here we have that word again .polar, polarity = direction}

 Apical dominance experiment

1 shows the apical bud

at the top.

In 2 and 3 the apical

bud is cut off.

Agar block without auxin does not prevent new lateral bud growth (#2). But either a
normal plant (without its tip removed) (#1) or a cut off plant covered with an agar
block containing auxin (#3), prevents lateral bud growth.

Thus, auxin (produced at the apex = top of the plant, or allowed to diffuse from this tip
into the agar block) plays a role in inhibiting lateral shoots from forming.

Many of you have done this experiment when you want your plants to be more bushy!
 Clearly auxin is made at the top of the plant. But it
seems to affect growth below the top..

Hormones are chemical signals that are produced in one part of an organism
and function in another

Auxin is made at the top of the plant and functions predominantly at the
bottom of the plant to stimulate root formation.

Is there a hormone to promote shoot and leaf formation?

Yes, cytokinin is made in the roots and is transported upwards to the top of
the plant to promote leaf and shoots!
Auxin made at the top of the plant, and transported downward (polar auxin
transport) is essential for root development
Cytokinin made at the bottom of the plant and transported upwards is
essential for shoot/leaf development
The amounts of auxin and cytokinin in different parts of the plant coordinate
how plants grow and what organs will form.

The plant is always sensing the ratio of auxin to cytokinin concentration and
vice versa.

When the cytokinin concentration is high and auxin concentration is low, shoots
and leaves form.

When the auxin concentration is high and cytokinin concentration is low, roots
form.
In the laboratory you can control plant morphogenesis (the development of different
forms) by just altering the amounts of auxin and cytokinin given to a lump of plant tissue.

No auxin High auxin High auxin Low auxin

No cytokinin High cytokinin Low cytokinin High cytokinin

Callus is a mass of undifferentiated (without form) cells

 What happens when the plant senses a high concentration of both auxin and cytokinin?

The plant cells do not differentiate, and remain undifferentiated. This is called a callus.

(In Lecture 13 we will learn about crown gall.)

In this slide the concentration is indicated by arrows where the tip of the arrow is the highest amount.
Now you can even better understand the apical dominance experiment

The plant senses the ratio of the amount (concentration) of cytokinin coming
up from the roots and auxin coming down from the top. When the source of
auxin is removed, by cutting off the top of the plant, the lateral buds will
sense that the amount of cytokinin is now higher (relative to the amount of
auxin) and they will start producing new shoots.
What happens when the both auxin to cytokinin are high?

In tissue culture in the lab, cells grow as a mass of undifferentiated

cells called a callus.
Plant hormones and totipotency..

A single plant cell can grow into a mass of undifferentiated cells in the
presence of auxin and cytokinin.

Totipotency is the ability to form an entire organism from a non-reproductive

cell. If you grow callus tissue from a single cell, you can then add hormones
to cause the cells of the callus to differentiate.
The callus tissue is then incubated on media containing only
cytokinin to induce shoots!

Do you need to add auxin for the roots to form ?

A. Yes
B. No
C. Dont know
Plants are Totipotent

Differentiation

De-differentiation
When a seed germinates which hormone is produced first, auxin or
cytokinin? (Imagine what would happen if either one is produced
first and then decide which is correct).

A. Auxin
B. Cytokinin
 What is the molecular basis for polar auxin movement?
What molecules in the cell control it?

2 different proteins are located in different positions in the cell.

Auxin (IAA) is taken up by influx carrier proteins (blue).

Auxin (IAA) is exported by efflux carrier proteins (red).

There are many more efflux proteins so whenever auxin (IAA) comes
into the cell it tends to be transported out and downward. Why?

Because efflux proteins are located at the bottom of the cell and
they will transport auxin downwards!
 FYI Auxin Chemistry

IAA (indole acetic acid) 2,4-D (dichlorophenoxyacetic acid)

Natural auxin Synthetic auxin

Synthetic auxins are used in abundance to kill plants !

Synthetic auxins are used as herbicides = to kill plants, as too

much auxin is not good.
 The powerful synthetic auxins 2,4-D and 2,4,5-T (=agent orange) were
used during the Vietnam war to defoliate the trees to expose enemies

Dioxin used in the chemical manufacture

of agent orange is the toxic factor!

Of 76 million Vietnamese, 1 million are disabled,

including 150,000 children!
Between 1962-71, U.S. helicopters sprayed 20 million gallons of
Agent Orange and other defoliants!
$60,000,000 required to clean up contamination. U.S. has contributed only $300,000 (0.5%) so far!

The above article was written around 15 years ago..

Now after over 40
years the US
finally has
stepped up!

Now will use

$43 million
to finally do a
clean up of still
contaminated
soils and
waterways.
How do small chemical messengers, hormones, cause such dramatic effects?

Phenotypes are the result of expression of particular genes at particular times

and in particular places.

How are genes turned on (activated) (expressed) ?

Whether or not a gene is expressed depends on whether or not it is
transcribed into mRNA!

To transcribe requires general enzymes such as RNA polymerase as

well as gene specific transcription factors.

FYI. auxin can be transported to the nucleus to activate

specific transcription factors that then have the ability to bind to
specific promoters to then stimulate transcription of genes that have
the ability to specifically respond to auxin!
What is causing the remarkable phenotype of this leaf?
Now that you know about photosynthesis and plant hormones you
can understand it!
The enormous floral heads of sunflowers do solar tracking!
Solar tracking requires that the stem bend.guess what hormone might
assist in differential bending due to cell expansion on one side of stem?
Auxin moves to the east side and then the west side of the
every day as part of the Circadian Clock =

Living organisms coordinate their biology and behavior

according to daily environmental changes in day-night cycle!

Humans have a clock that is important for regulating sleep

versus wakefulness.

Plants have a clock too.....what else might be regulated by

the clock in plants?

Photosynthesis, stomatal opening/closure, water uptake

Why do sunflowers go to the trouble of bending to the sun?

Facing the sun in the morning warms up the floral heads,

and this attracts pollinators!

Scientists tested this, by artificially warming the floral

heads that face away from the sunthen pollinators
also went to the dark side ;-)