21 Dec 2021

How to Tissue Culture Tomatoes?

Anjali Singh, MS

As a content and community manager, I leverage my expertise in plant biotechnology, passion for tissue culture, and writing skills to create compelling articles, simplifying intricate scientific concepts, and address your inquiries. As a dedicated science communicator, I strive to spark curiosity and foster a love for science in my audience.

Table of Contents

Introduction

Guess what is a fruit, but labeled as a vegetable?

Yes, tomatoes!

They are prepared and cooked like vegetables. And, also are the most popular flavoring agent when it comes to adding a tangy twist to different cuisines or served as a side sauce with many dishes.

Tomatoes are rich in nutrition, ranging from being a rich source of the antioxidant lycopene, vitamin c, potassium, to folate vitamin K. The fruit contains 95% water and 5% carbohydrates and fiber. And, knowing that it’s not surprising that some studies have found its use in reducing the risk of heart problems, cancer, and some other diseases.

This article talks about the process of cultivation of tomatoes using traditional techniques and its alternative advanced approach to produce the plants in thousands of numbers, fulfilling a huge population.

Get to know more about tomatoes!

It’s become easier to care about the plant when you know about it and when you understand its requirement based on its growth and appearance—because hey! Plants don’t talk!.

The scientific name given to tomato is “Solanum lycopersicum”. The plant was originally originated from Central America and South America and currently domesticated in many countries including India, Spain, China, Italy, Britain, and the Middle East, and North Africa. All these regions produce different varieties of tomatoes that differ in shape, size, and texture of the fruit and plant.

The plant typically grows 180 cm. But, the size and nature of the plant differ based on its variety, whether it's indeterminate or determinate type.

• Indeterminate type of tomatoes: They are also known as vining varieties. Their production tends to be more evenly spread throughout the season since they experience more leaf growth.

These varieties are needed to be staked and they are great for large gardens. Their uses in real life are Cherry and steak tomatoes that we use in our foods.

• Determinate type of tomatoes: They are also known as bush variety and grow 2-3 feet tall. A (relatively) short fruiting period and numerous ripe tomatoes are the main characteristics of these varieties and generally, they are more productive early in the growing season than vine varieties.

Furthermore, they do not need to be staked or caged and are very much ideal for small spaces and containers. The tomato paste that we usually use is a determinate variety that makes it ideal for cooking and canning.

Cultivation of Tomatoes

Conventionally, tomatoes are grown by using seeds. It is, however, not suitable for large-scale cultivation because it is time-consuming and costly due to the time and resources required for each breeding generation as well as difficulties with selecting the appropriate standards for cultivation.

Furthermore, tomato plants grown using conventional approaches are also prone to insect and pest attacks. And, due to the longer time requirements for cultivar development and existing biotic and abiotic stresses in normal field conditions, conventional seed propagation is less effective for introducing new cultivars and enhancing yields.

In vitro propagation techniques can be employed to enhance breeding efficiency and reduce the time period for cultivar development. Moreover, the technique can be used to improve tomato variety with better nutrients or resistance to diseases and biotic and abiotic stresses.

Tissue Culture of Tomatoes

Tissue culture is an advanced technique to grow disease-free, genetically altered, or high-valued tomato plants on a commercial scale.

The procedure mentioned below is taken from the study of Alatar et. al. (2017). Efficient and reproducible in vitro regeneration of Solanum lycopersicum and assessment genetic uniformity using flow cytometry and SPAR methods. https://doi.org/10.1016/j.sjbs.2017.03.008.

• Surface sterilization

• Collect mature seeds of the Solanum lycopersicum (of your desired cultivar) and thoroughly wash them under tap water for 30 minutes.
• Soak the seeds in 5% (v/v) liquid detergent solution for 5 min.
• Wash the seeds with sterile ultrapure water several times to remove all detergent residue.
• Surface sterilize the seeds using 2.5% (v/v) NaOCl (sodium hypochlorite) for 10 minutes.
• Then, immediately wash the seeds about 4 to 5 times with sterile ultrapure water.

• Seed germination

• Moisten cotton, placed in a magenta vessel with half-strength (½) Murashige and Skoog (MS) medium and transfer the sterile seeds on it.
• Before use, the media should be adjusted to 5.8 and autoclaved at 121 °C for 20 min.
• Incubate the culture seeds in dark for 48 hours and maintain under 50 µmol m–2 s–1 light provided by a cool white fluorescent lamp for a photoperiod of 16 h at 24 ± 2 °C in a growth chamber.

• Shoot initiation and Multiplication

• Excise cotyledonary leaf from the 8-day old aseptic seedling.
• Transfer the explant to MS medium supplemented with 6-benzyl adenine (BA; 5.0 µM), indole-3-butyric acid (IBA; 2.5 µM), and Kinetin (Kin; 10.0 µM).
• After every two weeks, transfer the cultures to a fresh medium.

• Rooting of the regenerated shoots

• Excise the in vitro regenerated shoots for about 4-5 cm in length.
• Transfer the shoots to ½MS medium supplemented with 0.5 µM indole-3-butyric acid (IBA).

• Acclimation

• After a few weeks, remove the plantlet from the cultured vessel and wash it gently under the tap water. This will remove all the adhered media to the root.
• Transfer the plantlets to pots containing sterile potting soil.
• For hardening, keep the potted plants in a growth chamber maintained at 24 ± 2 °C with diffuse light 50 µmol m–2 s–1 of 16/8 h photoperiod.
• For the next two weeks, irrigate the plants with half-strength MS salt solution.
• Then, after a month transfer the plants to the glasshouse under normal day length conditions.

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Happy Culturing!