How to Infect Tomato Plants with Pecto

The bacterium Pectobacterium carotovorum, commonly known as Pecto, is a major cause of bacterial wilt in tomato plants. Infected plants display symptoms such as yellowing and wilting of the foliage and stem collapse. Pecto mainly infects tomatoes through wounds or lesions caused by mechanical injury or insect feeding. This article will guide you on how to infect tomato plants with Pecto for research purposes.

Materials and equipment

Infecting tomato plants with Pecto requires several materials and equipment, including:

Tomato plants

Pectobacterium carotovorum culture

Agar plates

Sterile loop

Burner

Autoclave

Sterile syringe

Scalpel or razor blade

Preparation of Pecto culture

To infect tomato plants with Pecto, you first need to prepare a culture of the bacterium. Follow these steps:

Obtain a pure Pectobacterium carotovorum culture from a reliable source.

Prepare nutrient agar plates by dissolving agar powder in water and autoclaving at high pressure and temperature to sterilize.

Using a sterile loop, streak the Pecto culture onto the nutrient agar plates and incubate at 28°C for 24-48 hours until colonies appear.

Store the Pecto culture in a culture collection at -80°C for future use.

Infecting tomato plants with Pecto

Once you have prepared the Pecto culture, follow these steps to infect tomato plants:

Choose healthy tomato plants at the seedling stage and mark with a label.

Select a leaf on the tomato plant that is not damaged or diseased.

Using a scalpel or razor blade, create a small nick or wound on the leaf surface without damaging the underlying tissue.

Take a sterile syringe and collect a small volume (e.g., 20 μl) of the Pecto culture from the agar plate.

Inject the Pecto culture into the wound on the tomato plant leaf using the sterile syringe.

Repeat the process on different plants and control plants with sterile distilled water to conduct a comparative experiment.

Water the plants regularly and maintain at optimal growth conditions for the tomato plants.

Observe the tomato plants for symptoms of Pecto infection, such as wilting, yellowing, and stem collapse.

Conclusion

In conclusion, infecting tomato plants with Pecto requires preparation of a Pecto culture, selection of healthy tomato plants, and creating a wound on the leaf surface. The process involves the use of several materials and equipment, including sterile loops, syringes, and agar plates. Researchers can use the infected tomato plants to study the pathogenesis of Pecto and develop strategies for managing bacterial wilt in tomato crops.