22 Apr 2022

How to Tissue Culture The Hibiscus Plant?

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

Hibiscus is a flowering plant that belongs to the family Malvaceae. The family contains hundreds of plant species, native to tropical, subtropical, and temperate regions. Hibiscus is also known by the name rose mallow, hardy hibiscus, rose of Sharon, and tropical hibiscus. It’s a perennial and annual herbaceous plant, and also grows as woody shrubs or small trees.

About Hibiscus Plant

Hibiscus is a flowering plant that belongs to the family Malvaceae. The family contains hundreds of plant species, native to tropical, subtropical, and temperate regions.

Hibiscus is also known by the name rose mallow, hardy hibiscus, rose of Sharon, and tropical hibiscus. It’s a perennial and annual herbaceous plant, and also grows as woody shrubs or small trees.

The leaves of the plants are ovate to lanceolate in shape, alternate in arrangement and dentate on margins. Hibiscus flowers are large, trumpet-shaped with 5 or more petals, and available in a variety of colors, ranging from white to pink, red, blue, orange, peach, yellow or purple. The fruits of the plant are five-lobed capsules and possess many seeds in each lobe.

The plant is used for a spectrum of purposes, including landscaping, paper-making, rope-making and construction, beverage and food in some countries, such as Mexico, and in folk medicine.

In this article, we will cover the tissue culture technique of the hibiscus using nodal cuttings as explants.

Micropropagation of Hibiscus

In addition to being used as a medicinal plant, the Hibiscus plant is also predominantly used for landscaping different purposes.

Conventionally, the plant is grown using seeds or stem cutting. However, the growth rate of plants grown using these methods is considerably slow. And, the other challenge is to grow a huge number of mother plants to grow plants through cutting. In such cases, clonal propagation or tissue culture is an efficient technique to overcome the challenges.

In tissue culture, small tissue sections are obtained from the plants and cultured on a media in lab condition, mimicking the natural condition of the plants. The advantages of the technique include:

• High-reproduction rate of plants.
• Production of healthy and disease-free plants.
• Large-scale production of plants in small areas.
• Year-round production of plants, irrespective of their growing season.
• Allow the conservation of plant germplasm.

However, the growth and development of plants in lab conditions depend on several factors, including:

• Macro-and microelement composition
• Vitamin mixture
• Sugar concentration
• Growth regulator composition
• Climatic condition

Procedure to Tissue Culture Hibiscus

Here’s the procedure to tissue culture Hibiscus rosa-Sinensis using nodal cuttings. The protocol is taken from the study of Christensen, Brian & Sriskandarajah, Sridevy & Serek, M. & Müller, Renate. (2008). In vitro culture of Hibiscus rosa-Sinensis L.: Influence of iron, calcium, and BAP on establishment and multiplication. Plant Cell, Tissue, and Organ Culture. 93. 151-161. 10.1007/s11240-008-9354-4..

Plant Material and Surface Sterilization

• Take the nodal cuttings without leaves from the mother plant.
• Sterilize the nodal cuttings n 1.5% (v/v) NaOCl and 0.03% (v/v) Tween 20 for 20 minutes.
• Rinse the cuttings with three changes of distilled water.

Establishment

• Prepare a basic MS medium, consisting of MS macro- and microelement, 30 g l-1sucrose, 100 mg l-1 Myo-inositol, 3 g l-1 Gelrite, and 0.5 g l-1 MES.
• Adjust the pH of the media to 6.3 and then autoclave it at 121°C and 103.5 kPa for 20 min.
• Add 197 µM Fe-EDDHA, 9 mM calcium ions as CaCl2H2O, and filter sterilized 2.2 µM BAP to the prepared media.
• Take the surface-sterilized nodal cuttings and place them on the media.
• Maintain the cultures in a growth room at 24°C and 16 h-photoperiod of 45 µmol m-2s-1(PAR) provided by cool-white fluorescent tubes.

Subculturing

• Subculture the shoots from establishment media on the same media as mentioned above (modified MS medium containing 2.2 µM BAP and concentrations of calcium at 9 mM and iron at 295 µM provided as Fe-EDDHA).
• Subculture the obtained shoots again in the same media.
• Maintain the cultures in a growth room at 24°C and 16 h-photoperiod of 45 µmol m-2s-1(PAR) provided by cool-white fluorescent tubes.

Root Induction

• Take the shoots from the second subculture and place them in the root induction media, which is a half-strength modified MS medium containing 2.7 µM NAA.
• Maintain the cultures in a growth room at 24°C and 16 h-photoperiod of 45 µmol m-2s-1(PAR) provided by cool-white fluorescent tubes.

Acclimatization

• Sterilize the peat by autoclaving it at 121°C and 103.5 kPa for 20 min.
• Place the plants into a clear plastic container containing sterilized peat.
• Keep the plantlets in a greenhouse, at 22°C/20°C day/night, and 70% RH, until flowering.
• Irrigate the plants once a day with a standard fertilizer.

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So, visit plantcelltechnology.com today and find out more about our product and services and how they help you to excel in your tissue culture processes.

Happy Culturing!!