04 Lab - Asexual propogation and tropisms WQ2024 (1)

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Feb 20, 2024

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Name / student ID Date: PLS002 WQ2024 – Lab 04 1 Objectives Many important agricultural crops are propagated asexually, i.e., vegetative propagation. In essence, asexual propagation is the production of a whole plant from a small portion of another plant (this is only possible because all living plant cells contain the full genome of the plant – totipotency). Every plant produced by vegetative propagation is genetically identical to the parent plant, i.e., it is a clone. The main objective of this lab session is to familiarize you with the theory and practice involved in plant asexual propagation ( exercises 2, 3 and 4 ). We also look at gravitropism and phototropism in exercise 1 - so you can see the response of a plant to environmental stimuli. Lab 03 – Assignments check list Pollination of tomato and fruit development with and without fertilization – For Assignment 8 data. There are no questions to answer and no points available this week, but you need to check your plants and repeat the pollination treatments. Exercise 1 Tropisms – Look at the demonstrations with your group and four questions to answer here. Exercise 2 Grafting – after the in person demonstration do your vine graft and answer three questions. Exercise 3 Cuttings – make your cuttings as described in this schedule and answer two questions Exercise 4 Tissue culture – Look at the samples provided and watch the video then answer two questions. Cucumber growth (For Assignment 10 ) You need to record the plant height, tie up your growing plant and harvest and record any fruit – number of fruit and total fruit weight. Health and safety guidance Take care not to cut yourself with the pruners, scissors and grafting tools used in this lab. Also remember the standard safety issues in glasshouses: Wet floors - trips and slips, Fans and lights – hair tied back Benches (e.g. fingers between moving benches) and other snags Electric outlets – do not get these wet Look out and take care of yourself. Label, label, LABEL! Make sure you include your section number, name/group number, date/lab number and the pot contents on the labels. Good organization, including labeling, is important in science LAB 04 PLANT PROPAGATION (GRAFTING, CUTTINGS); TROPISMS
Name / student ID Date: PLS002 WQ2024 – Lab 04 2 methodology and is the only way you will keep track of your plants. Exercise 1: Tropisms: plant response to gravity and light To illustrate the effect that light and gravity can have on plant growth, and how plants response to these various signals, tomato plants have been initially grown in pots under “standard” conditions (in greenhouse), and then placed in different conditions, or “treatments”, as follows: - In the glasshouse, one tomato plant remained upright and the other pot was tilted over on its side. - In a closed box with a hole cut in the south side, one tomato plant remained upright and the other pot was tilted over on its side. - In a tent or box without any light (complete darkness), one tomato plant remained upright and the other pot was tilted over on its side. After looking at these demonstrations in the glasshouse answer the questions below: Gravitropism 1) What happened to the plant that were tilted on its side in the dark? In particular give the rough angle that may have formed in the younger section of the stems and note which direction the stem grew in the dark. Why are the plants placed in the dark the best indication of the gravitropic response? 2) - How can bending on a stem occur in response to gravity? (Think about how a plant might “sense” gravity and alter stem growth – cell number, cell length or both & on which side of the stem). Phototropism 3) What happened to the plants that were put in different light conditions (scattered light in the glasshouse, one-directional light, no light)? Was the response different if the plant was put on its sides? In particular, note the angle that may have formed in the younger section of the stem, and whether the leaves also reoriented. 4) What is the difference between phototropism and photoperiodism (see lecture 6 PDF)? Exercise 2: Grafting Watched the demonstration of cleft grafting using parts from different grape varieties (one, the rootstock, the other, the scion) . Grafting is often performed in the winter or early spring, when both the rootstock and the scion are dormant – this type of grafting is called “bench grafting”, because it is
Name / student ID Date: PLS002 WQ2024 – Lab 04 3 done at the bench (as opposed to in the field). For more information on grafting, you are encouraged to check out this excellent resource by NC State University Extension: https://content.ces.ncsu.edu/grafting-and-budding-nursery-crop-plants (A copy of grafting and budding nursery crop plants is also in the Canvas Files area). Now make your own vine plant by grafting and then answer the following questions: 1. Can monocot plants be grafted? Why or why not? 2. Why is it important to align the cambium of the rootstock with the cambium of the scion? 3. Name two other types of grafting (other than cleft grafting) that are commonly used on fruit trees Exercise 3: Cuttings Watch the demonstration of how to take cuttings . A number of plants will be provided for you to take cuttings from. We hope to have cinnamon scented geranium ( Pelargonium limoneum ), purple heart ( Tradescantia pallida ), trailing Coleus ( Lava rose ) and ornamental fig ( Ficus benjamina) for you to use. These are all great house plants. Please make cuttings of all the species provided and try with and without rooting hormone (indole-3-butyric acid, an auxin) in order to observe the effects of the rooting hormone in a few weeks time. You need to label your plants and treatments and to keep them separate. Procedure Prepare pot labels with your name, section number, group number, treatment (control or hormone), plant information (type of plant), date and lab number (= Lab 03). 1. Cut two stem tip cuttings from each of the plants provided, with pruning shears. 2. One cutting of each plant type will be dipped in rooting hormone before being stuck in rooting media. 3. We will provide flats for each of the types of cuttings. Place your plants in pots in the correct flat: - Plant A cutting dipped in hormone - Plant A cutting without hormone (= control) - Plant B cutting dipped in hormone
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Name / student ID Date: PLS002 WQ2024 – Lab 04 4 - Plant B cutting without hormone (= control) 4. At the end of the quarter we will look at the amount of rooting in each treatment and, we hope you will be able to take their own propagated plants back home. After doing your cuttings answer the following questions: 1. Which “treatment” do you expect to be most successful at obtaining a new plant from the cutting? Explain. 2. Why is it crucial to keep the cuttings clean and moist? Exercise 4: Tissue culture observation Each living cell in a plant contains the same genetic material as all the others, and has the potential to form a complete plant (i.e., each cell is totipotent). “Plant tissue culture” is the practice of growing entire plants from generally very small plant parts (such as meristems, pieces of leaves, nodal sections of stems, even anthers). As we have discussed in the lectures bulking up plant tissue in aseptic conditions, generating meristems or embryos (somatic embryogenesis) from this bulked-up material and then taking them through to be individual plants can be very important commercially and in research programs. Have a close look at the lettuce tissue culture plates which are provided. We have tried to get plates which show the different stages of the lettuce tissue culture procedure. Watch the tissue culture tutorial at https://www.youtube.com/watch?v=uPuxS1kxdVY (you will see that there are a number of similar videos if you’re interested) and answer the following questions. 1. What are the main advantage of tissue culture which explain why it is so extensively used in the horticultural trade? 2. Somaclonal variation is the variation seen occasionally in plants that have been produced by plant tissue culture (particularly if plants are produced from callus rather than from meristems) . How do you think this might happen? Explain one advantage and one disadvantage related to this phenomenon.

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