Give a five steps procedure on how to collect the phloem exudates using aphids (short steps)
1. Identify a suitable plant: Choose a plant species that is known to have a high phloem sap flow and can easily be infested by aphids.
2. Create an aphid colony: Establish a colony of aphids by collecting a group of aphids from their natural habitat or purchasing them from a reliable source.
3. Infest the chosen plant: Place the aphids on the selected plant and allow them to settle and establish themselves by feeding on the phloem sap.
4. Collect the phloem exudates: Set up a suitable apparatus, such as a glass capillary or silicon tubing, to pierce or cut a phloem sieve element. Position the apparatus close to the aphids and place a collection solution at the other end to collect the phloem exudates.
5. Monitor and collect the exudates: Monitor the collection system regularly and collect the phloem exudates that have been secreted by aphids feeding on the plant. Store the collected exudates appropriately for further analysis or experimentation.
Step 1: Obtain healthy aphids - Collect healthy aphids by carefully picking them from a plant that is known to have active sap flow and a good number of aphids.
Step 2: Set up a suitable environment - Transfer the aphids to a container or a glass slide, keeping them in a controlled environment such as a glass petri dish with a moist cotton pad or a plant stem immersed in water.
Step 3: Wait for exudation - Allow some time for the aphids to acclimate to their new surroundings. The aphids will start to feed on the plant's phloem sap, and after a while, they will excrete the excess sap as honeydew.
Step 4: Position a collecting medium - Position a collection surface, such as a clean glass microscope slide or a piece of wax paper, underneath the aphids to collect the phloem exudates as they feed.
Step 5: Collect the exudates - Gently touch the surface of the exudates with a dropper or a capillary tube to collect the phloem exudates. Be careful not to disturb the feeding aphids or contaminate the collected exudates.
Note: It is important to handle the aphids gently to avoid injuring them and to ensure proper feeding. Additionally, depending on the experiment or analysis, the collected phloem exudates might need to be processed or stored a certain way for further use.
Sure! Here's a five-step procedure for collecting phloem exudates using aphids:
Step 1: Prepare the aphids
- Obtain a population of aphids. You can either collect aphids from nature or purchase them from a supplier.
- Make sure the aphids are healthy and fed by rearing them on a suitable host plant. Common species used for this purpose include green peach aphids (Myzus persicae) or cabbage aphids (Brevicoryne brassicae).
- Rear the aphids in controlled conditions, providing them with a continuous supply of fresh plant material to feed on.
Step 2: Isolate the aphid
- Select a healthy aphid from your population, preferably a mature adult.
- Using a soft paintbrush or a fine-tipped forceps, gently detach the aphid from the plant to isolate it.
- Be careful not to harm the aphid, as it needs to remain intact for the following steps.
Step 3: Position the aphid
- Place the isolated aphid on a small, clear plastic or glass microscope slide.
- Ensure that the aphid is positioned upright, with its mouthparts (stylets) in contact with the slide's surface.
- Allow the aphid to acclimate to the slide for a short period, as this can enhance the phloem exudate collection.
Step 4: Collect the exudate
- Using a high-resolution microscope or a stereomicroscope, carefully observe the aphid's stylets under magnification.
- Look for the appearance of clear, watery droplets (phloem exudates) forming at the tip of the aphid's stylets.
- With the aid of a fine microneedle, capillary tube, or glass tube, gently and skillfully touch the exudate droplets to collect them.
- Be cautious not to disturb the aphid or cause any damage during the collection process.
Step 5: Store and analyze the exudate
- Transfer the collected phloem exudates into a suitable container, such as a microcentrifuge tube or a vial.
- Seal the container to prevent evaporation and contamination.
- Store the exudates at appropriate temperatures (commonly refrigerated) to maintain their integrity until further analysis or experimentation.
Remember, collecting phloem exudates using aphids requires patience, precision, and attention to detail. It is crucial to handle the aphids and equipment with care to obtain reliable results.