Bagaimana Infeksi Mikoriza pada Akar?

Summary

This tutorial guides students through the process of observing mycorrhizal root colonization using a staining method. It covers the preparation and treatment of root samples, including washing, soaking in KOH, HCl, and dye, followed by mounting and microscopic examination. The key mycorrhizal structures—vesicles, arbuscules, hyphae, and spores—are identified. The tutorial also demonstrates how to quantify root colonization using a percentage formula and categorize the infection level into various classes, providing a clear and educational approach for analyzing mycorrhizal colonization in plant roots.

Takeaways

• 😀 The tutorial demonstrates how to observe mycorrhizal colonization in plant roots using a staining method under a microscope.
• 😀 The primary tools and materials needed for the procedure include KOH solution, HCl, Trivago dye, petri dishes, and dissecting tools.
• 😀 The first step involves cleaning plant roots to remove soil and debris, using a sieve to prevent fine roots from being discarded.
• 😀 Roots are then cut into small pieces (approximately 2 cm) and placed into test tubes for further processing.
• 😀 The roots are treated with a 10% KOH solution to soften the roots and extract cytoplasmic contents for better observation.
• 😀 After heating the roots in KOH solution at 80°C for 2 hours, they are washed several times with clean water to remove excess solution.
• 😀 The next step involves immersing the roots in 2% HCl solution at 80°C for 2 minutes, which helps to prepare the roots for staining.
• 😀 Roots are then soaked in 0.05% Trivago dye for 24 hours to ensure thorough staining of the mycorrhizal structures.
• 😀 After staining, roots are washed again with distilled water and placed on a microscope slide for observation under varying magnifications.
• 😀 Key mycorrhizal structures to look for under the microscope include vesicles (oval-shaped), arbuscules (tree-like structures), hyphae (thin thread-like structures), and spores (round and perfect in shape).
• 😀 The percentage of mycorrhizal colonization is quantified using a formula based on the number of infected roots versus total observed roots, which is then classified into different categories based on colonization percentages.

Q & A

What is the purpose of the root colonization observation?

The purpose is to observe the development of mycorrhizae infecting the roots of plants using a staining method under a microscope.

What materials are needed for the root colonization observation?

The materials needed include Koh 10% KOH solution, 2% HCl solution, 0.05% Trivago stain, glass slides, cover glass, test tubes, petri dishes, tweezers, pipettes, film files, and plant root samples.

What is the function of the 10% KOH solution in the procedure?

The 10% KOH solution is used to release the cytoplasm and soften the roots, making them easier for further observation and staining.

Why is HCl solution used in the procedure?

The 2% HCl solution is used to fix the roots and make the staining process easier.

What does the 0.05% Trivago stain do in this experiment?

The Trivago stain is used to color the root samples to make the mycorrhizae, including vesicles, arbuscules, hyphae, and spores, visible under the microscope.

How long should the root samples be soaked in Trivago stain?

The root samples should be soaked for 24 hours to ensure the stain is fully absorbed.

What are vesicles and what is their function in the mycorrhizal infection?

Vesicles are oval-shaped structures that store nutrients and are less than one millimeter in diameter.

What role do arbuscules play in mycorrhizal infection?

Arbuscules, which resemble tree branches, are the sites of nutrient exchange between the plant and the mycorrhizae, helping the plant to absorb essential nutrients.

What is the purpose of using cover glass in the preparation of the root sample?

The cover glass is used to reduce shifting of the root sample and protect it during observation under the microscope.

How is the percentage of root colonization calculated?

The percentage of root colonization is calculated using the formula: (number of infected roots / total number of observed roots) * 100%.