14 Oct 2021

How to Tissue Culture Anubias?

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

Do you have an aquarium at your house or you are into the aquarium business? Well, in either case, plants have been a great element to decorate the aquarium, make it alive, and give fishes the feeling of their natural habitat. To give them a cozy environment to play and wander. The natural aquarium plants provide shelter to your fish and a place to breed and take care of their young.

Further, the presence of natural plants in aquariums facilitate the absorption of CO2 and ammonia (released fish waste) from water and release oxygen, required for the growth and survival of the inhabitants of your aquarium.

Aquarium plants have become popular with the increasing demands of aquariums by people for their houses or business plants. The commercial need for these plants has attracted many culturists worldwide to try their expertise in curating techniques to grow these plants on a large scale.

Some of the well-known plants are Anubias, moneywort, hornwort, tiger lotus, Hygrophila polysperma, duckweed, and water wisteria.

This article covers the method for in vitro culturing of Anubis and the suitable environment supporting their growth.

About Anubias

Anubias is a craze for hobbyists and culturists worldwide. They are well-known for their hardy nature (can be kept in a variety of conditions) and beautiful green leaves and are easy to introduce in your aquarium. They keep the tank water clean and oxygenated.

Anubias genus belongs to the family of Arecaceae. It contains aquatic and semi-aquatic flowering plants. The plants are native to tropical central and western Africa. They are mainly found in the habitat of rivers and streams, but can also be found in marshes.

Anubias has broad, thick, dark leaves that come in many different forms. Their maximum height is around 7.5 inches, but it can vary depending on the surrounding conditions.

Image: The Anubia plant.

The plants are slow in growth, thus require less maintenance, except a bit of trimming now and then. To maintain these plants you need to follow the measure explained below:

Keep your tank clean regularly to remove pollutants that eventually become toxic and kill any life in your tank.
Add nutrients supplements whenever you start observing the slow growth in plants by ensuring that you are not adding too much a nutrient.

Tissue Culture of Anubias barteri var. Nana

Anubias barteri is the most commonly grown plant in aquariums. It has several varieties including Anubias barteri var. Barteri, A. barteri var. Angustifolia, A. barteri var. Caladiifolia, A.barteri var. glabra, and A. barteri var. Nana. Among all these A. barteri var. Nana is mostly preferred for aquariums.

Anubias doesn’t require meticulous maintenance of the plants. You only simply need to trim back the leaves near the rhizome with a sharp pair of scissors. They prefer the pH of water in the range of 6.5 to 7.5, but it can also adapt to a pH value from 5.5 to 9.0 because of its extreme pH tolerance nature. The perfect temperature to maintain these plants is between 22 to 27°C (72-82°F).

The conventional technique to grow Anubias is by using stolon division. But, it has a very slow multiplication rate, thus, an inefficient technique for the commercial propagation of plants. Micropropagation of Anubias is an amazing alternative technique to grow plants on a large scale with better and higher output.

Here, a protocol to culture A. barteri var. Nana using shoot tip culture is mentioned, taken from the study of Kanchanapoom, K., Chunui, P., & Kanchanapoom, K. (2012). Micropropagation of Anubias barteri var. Nana from Shoot Tip Culture and the Analysis of Ploidy Stability. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 40(2), 148. doi:10.15835/nbha4027520.

Procedure

Collect the young plantlets of A. barteri var. Nana.
Surface sterilize it using 0.5% (w/v) mercuric chloride solution containing 2 drops of Tween-20 emulsifier per 100 ml solution for 3 min.
Wash the treated plants three times with sterile distilled water to remove any remnants of disinfectants.
Then, surface sterilizes the explants using a dilution of 10% (v/v) commercial Clorox™ which yields 5.25% NaOCl and 2 drops of Tween 20 per 100 mL solution for 15 min, followed by 5% (v/v) Clorox™ for 5 min.
Rinse explants three times using sterile distilled water.
Excise 3.5 mm shoot tip explants before culturing them on a basal medium containing 3% sucrose for their growth.
After six weeks, transfer small shoots with a pair of leaves to MS medium supplemented with 3 mg/L BA and containing 3% sucrose and 0.82% agar.
Adjust the pH of the media to 5.8 before sterilizing at 121 ℃ for 20 minutes.
Maintain the cultures at 25±1˚C in a culture room with a 16-h light photoperiod.
Subculture the explants at the interval of eight weeks.
Maintain the cultures at 25±1˚C in a culture
room with a 16/8 h light/dark photoperiod under the illumination of 20 µmolm-2 s-1 photosynthetic photon flux intensity provided by cool white fluorescent light.
Rooting can be regenerated by transferring the shoots to MS media either devoid of any plant growth regulators or kinetin singly.

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Happy Culturing!!