GREEN ALGAE DIVISION CHLOROPHYTA

Introduction Of the nearly 5,500 species of green algae, approximately 90% are restricted to the freshwater environment: damp soil, rivers, lakes, ponds, puddles, tree bark, and even the hair of polar bears. The marine representatives are limited to relatively few orders and are common in the intertidal and upper subtidal regions. Like vascular plants, green algae have chlorophylls a and b in addition to a variety of carotenes and xanthophylls that act as accessory pigments. Nutrition is autotrophic, with the reserve carbohydrates stored in plastids in the form of starch. Green algae exhibit a wide variety of thallus forms, ranging from single cells to filaments to parenchymatous thalli. In tropical and subtropical waters, many forms may be calcified. Reproduction occurs asexually by fragmentation or by the production of spores that develop directly into new individuals, or sexually by the union of two gametes. While the sexual gametes and asexual spores may look very similar, they can be differentiated by the number of flagella; sexual gametes have two flagella and asexual spores have four flagella. In many species the entire thallus becomes reproductive, a term called holocarpic. Systematics A few distinguishing characteristics separating key orders for marine Chlorophyta (Class Ulvophyceae) are below (from Van de Hoek, page 95).
Order Thallus Chloroplasts Reproduction & Life History (asexual; sexual) Examples

Ulvales Cladophorales Caulerpales

parenchymatous

single, parietal, several pyrenoids parietal, reticulate, few to many pyrenoids numerous, discoid, w/o pyrenoids

coenocytic cells united end-end much branched, coenocytic, nonseptate

zoospores; gametes iso/ anisogomous. mostly isomorphic stages fragmentation, zoospores; biflagellate gametes none; biflagellate gametes

Ulva

Cladophora, Chaetomorpha, Acrosiphonia Codium, Bryopsis

Morphology The green algae are well represented in the marine plankton and damp terrestrial environments, with many species occurring as unicellular organisms. These include motile vegetative cells that produce vegetative spores and/or gametes (e.g. Chlamydomonas), non-motile vegetative cells that produce flagellated reproductive structures (spores/gametes) (e.g. Chlorococcum), and lastly non-motile vegetative cells without motile reproductive structures (e.g. Chlorella). Colonial green algae (e.g. Volvox) are not commonly encountered in the marine environment, though they are common residents of soil and standing freshwater. Colonies (synobia) are made up of distinct single cells held together by mucilage.

A giant evolutionary step was made in the green algae when the cells that resulted from a mitotic event remained contiguous. Connected cells enabled both multicellularity and the complex functions this required. Filamentous green algae are well represented in the sea either as unbranched, simple uniseriate (single thread) filaments of cells that show little or no thallus differentiation (e.g. Chaetomorpha, Ulothrix), or as branched filaments (e.g. Cladophora). If a single cell divides in only one plane, a filament is formed. However, if that single cell divides and remains adherent in two or three planes, a parenchymatous multicellular thallus is formed (e.g. Ulva, Enteromorpha). A variation of the filamentous thallus that is tubular and lacks cross walls (except when delimiting reproductive structures) is called siphonous. The siphonous filaments may occur singly (uniaxial, e.g. Bryopsis) or may be aggregated into a pseudoparenchymatous multiaxial system (e.g. Codium). Most of the siphonous marine green algae are tropical, but a few genera are prominent in the flora off the west coast of North America (e.g. Codium).

Objectives In todays lab, you will examine in detail marine green algae that represent the wide range of morphologies and reproductive strategies. The green algae occur as unicellular and multicellular thalli. Within the unicellular and colonial forms, some are motile and produce vegetative cells and/or gametes (e.g. Chlamydomonas), some are not motile as vegetative cells but produce flagellated spores/gametes (e.g. Chloroccum), and some are not motile as either vegetative or reproductive cells (e.g. Chlorella). Within the multicellular forms, some are filamentous (e.g. Chaetomorpha), some are parenchymatous (e.g. Ulva), and some are pseudoparenchymatous (e.g. Codium). You will be presented with several fresh and prepared examples of each form. Examine representatives from each group and make careful drawings of the key features that separate them. Notebook Requirements (14 drawings) 1) Chlamydomonas- drawings of different life stages (at least 2) 2) Volvox- drawings of different steps in life cycle (at least 2) 3) Chaetomorpha- 2 drawings (thallus & single cell) 4) Cladophora- 2 drawings (thallus & single cell) 5) Ulva- 2 drawings (thallus & cross section) 6) Codium and Bryopsis -2 drawings (thallus & cross section) a) Codium cross section on prepared slide b) Bryopsis thallus on compound scope c) Bryopsis key out as known 7) Unknown 1- 1 drawing & steps to key out 8) Unknown 2-1 drawing & steps to key out

A. Unicellular Forms
Key Organism: Chlamydomonas (Class Chlorophyceae, Order Volvocales)

Unicellular forms can be motile or non-motile. Chlamydomonas is a single-celled motile alga commonly found in damp soil, ditches, tide pools and occasionally in salt marshes. The organism is typically egg-shaped and has a large cup-shaped chloroplast containing a proteinacious bodythe
2

pyrenoid, which functions in starch polymerization. The nucleus is often difficult to see because of the prominent chloroplast. 1. Examine mating strains of Chlamydomonas. Identify the steps in the life cycle. Draw 2 different life stages in detail. Be sure to draw and label chloroplast.

B. Colonial Forms
Key Organism: Volvox (Class Chlorophyceae, Order Volvocales)

While colonial green algae are not common in the marine environment, they are widespread in soil and standing fresh water. Some are motile flagellated species (e.g. Gonium, Pandorina, Eudorina, Volvox), while others are non-motile colonies that produce flagellated reproductive cells (e.g. Chlorococcum). 1. Identify the different colony types of Volvox. Draw at least 2 different colony types. C. Filamentous Forms
Key Organism: Chaetomorpha (Class Ulvophyceae, Order Cladophorales)

Chaetomorpha is a very common unbranched filamentous alga in the intertidal areas of central California. It is usually attached to pier pilings or to rocks and looks like green hair.

Chaetomorpha lineata

1. Examine the Chaetomorpha thallus under the compound microscope. Draw a single cell and compare its inclusions to those seen in Chlamydomonas. How does the chloroplast shape differ? Is there an eyespot? Draw the thallus, note the holdfast if present. 2. If we have reproductive material, we will set up a demonstration in which we cause the thalli to become reproductive. To do this, place a piece of the thallus under a light source and observe under a dissecting scope. As the thallus heats up, it will begin releasing all the pigmented material inside. What is the term for this? Can you think of a benefit of reproducing this way?
Key Organism: Cladophora (Class Ulvophyceae, Order Cladophorales)

Cladophora is a branched filamentous alga, growing commonly in both fresh and salt water.

Cladophora columbiana

Cladophora is found growing attached to rocks and other objects by means of a specialized cell, the holdfast. 1. Note and draw the holdfast if it is present. Draw the extensive branching. 2. Observe the chloroplast in living cells of Cladophora. How does this chloroplast shape differ form the chloroplast in Chlamydomonas and Chaetomorpha? 3. IKI stain reacts with starch, turning it dark in color when the reaction is positive. Stain a sample with IKI stain. Is the reaction positive or negative? What does this tell you? D. Parenchymatous Forms
Key Organisms: Ulva (Class Ulvophyceae, Order Ulvales)

Ulva spp. are common and widespread throughout the world.

Ulva intestinalis

Ulva californica

1.

Observe the morphologies of these two species with the naked eye and under the dissecting scope. Where does growth appear to occur? Is it diffuse, or localized to a meristem? Look to see what structure anchors these algae to the substrate. Make a thin cross sections of Ulva and draw the internal details. How many cells thick are they? Draw the individual cells. What is the shape of the chloroplast? Is an eyespot present? Is the chloroplast facing the inner or the outer part of the thallus? How many chloroplasts are there per cell? If we have reproductive material, we will set up a demonstration in which we cause the thalli to release gametes. The gametes are positively phototactic, whereas the zygotes and zoospores are negatively phototactic. Can you hypothesize a reason for this? For ideas, look over the diagram of Ulvas life history at the end of this lab.

2.

3.

E. Siphonous Forms
Key Organisms: Codium, Bryopsis, (Class Ulvophyceae, Order Caulerpales). Note, we do not have a fresh sample of Codium. We have prepared slides for you to use.

Codium setchellii

Codium fragile

1. Siphonous algae are multinucleate and tubular, with few transverse septa (cell walls). Observe and draw the overall external morphology of the samples. Codium (Order Codiales) is a siphonous alga in which the siphons are entwined together to form a spongy, pseudoparenchymatous plant. Examine and draw the external morphology of Codium. 2. Squash a small piece of Codium between two microscope slides and observe. Are there chloroplasts in these filaments? Do you see cross walls? Observe the tip of the thallus. Are cross walls present? Plastids? What is the shape of the plastid? Do you see any reproductive structures? Look over the diagram of the life history and close-up showing utricles. Can you suggest a purpose for these? Bryopsis Draw the thallus using a compound scope. Key this algae out as our known species.

3. 4.

F. Identifying Algae to Species To practice keying out algae, you will need to key individual samples to species presented to you as knowns. The species names will be given to you, but you will need to work through the dichotomous key in the Marine Algae of California to key them out. Once you have finished keying the known species, choose one of the unknowns and try to key it to species. If time permits, do this for a second unknown. Make careful drawings of anything you think might be relevant. For all species you key out, it is important to write down the steps you take as you work through the key. This way, if you are incorrect, you can retrace your steps much more quickly and find where you went wrong.

Life Cycle of the Cladophorales

Isomorphic alternation of generations e.g. Cladophora columbiana
meiosis occurs in apical cells

1N zoospore

2N filamentous sporophyte

1N filamentous gametophytes

1N gametes

2N zygote

syngamy

Life Cycle of the Caulerpales

Animal-like (diplontic) e.g. Codium fragile
syngamy

1N gametes

meiosis occurs in utricles 2N adult

2N zygote

Life Cycle of the Ulotrichales

Haplontic e.g. Ulothrix

syngamy

1N gametes

2N zygote meiosis occurs in zygote

1N gametophytes

1N codiolum stage

1N zoospores

Life Cycle of the Ulvales

Isomorphic alternation of generations e.g. Ulva lactuca
syngamy

1N gametes 2N zygote

1N gametophytes

2N sporophyte meiosis occurs in cells of sporophyte

1N zoospores