Thin Cell Layer Culture of Dendrobium sp. and Cymbidium sp.

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Thin Cell Layer Culture of Dendrobium sp. and Cymbidium sp.

Tran Van Minh1,2*
1International University, Vietnam National University Ho Chi Minh City, Vietnam
2Key Lab of Plant Cell Biotechnology, Institute of Tropical Biology, Vietnam
*Corresponding author
DOI: https://doi.org/10.51584/IJRIAS.2023.8611
Received: 29 May 2023; Revised: 19 June 2023; Accepted: 26 June 2023; Published: 07 July 2023

Abstract: Vietnam, a nation located in tropical region, has an important plan for flower development esspecially in tropical orchid; and the first barrier is seedling production. Some orchid species are difficult to regenerate, and thin cell layer is a method for manipulation. Dendrobium in low land tropic and Cymbidium in high land tropic were used as model to produce seedlings for high demand from farmers. Dendrobium sp.: Young shoot from the pot was used as materials for in vitro shoot regeneration on the medium MS + BA (1mg/l) + IBA (0.1mg/l) after 30 days and the in vitro shoots were cultured on the same media for multiplication. The shoots were prepared separately to leaves and shoot tip that were cultured on the thin cell layer culture medium MS + 2iP (1mg/l) + IBA (0.1mg/l) to raise 6-8 shoots/shoot tip sample and 4-6 PLB/leaf sample. Diversity of gene in cultivation was not recorded in difference about PLB initiation with 4-6 PLB/sample. Shoots and PLB were multiplied on the medium of MS + Adenine sulfate (10mg/l) + IBA (0,1mg/l) + BA (1mg/l). Shoots were rooted well on the medium MS + IBA (1mg/l). The thin cell layer culture of Dendrobium sp. was established. Cymbidium sp.: Young shoot from the pot was used as materials for in vitro shoot regeneration on the medium MS + peptone (1g/l) + BA (1mg/l) + NAA (0.5mg/l) after 30 days. In vitro shoots were used as materials and were multiplied on the medium MS + peptone (1g/l) + BA (1mg/l) + NAA (0.5mg/l). Shoots were released separately to leaf and shoot tip and cultured on the thin cell layer culture medium MS + peptone (1g/l) + BA (1mg/l) + NAA (0.5mg/l) to raise 4-6 PLB/shoot tip sample and 3-5 PLB/leaf sample. PLB raised from shoot tip was bigger than from leaf. PLBs were multiplied on the medium of MS + BA (1mg/l) + NAA (0.5mg/l). PLBs were regenerated and rooted on the medium MS + BA (0,1mg/l) + rhizogen (5mg/l) with high rate of 100% after 45 days. The diversity of gene was recognized that it was not different on 5 clones cultured about PLB initiation with 4-6 PLB/sample. The thin cell layer culture of Cymbidium sp. was established. Application of TCL method for two orchid sepcies leading to create primary materials as multiple shoots and PLB for far micropropagation.

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Keywords: Dendrobium sp., Cymbidium sp., thin cell layer, shoot regeneration

I. Introduction

Mineral composition and plant growth regulators play an important role in the growth and development of plants, especially in vitro plants [1]. Mineral nutrient composition in each environment has different nutrient composition and concentration. Each plant species is suitable for a certain composition and concentration of mineral nutrients. Plant growth regulators are commonly used in vitro to regulate homeostasis and control morphogenesis and organogenesis in vitro [2]. The balance of growth regulators determines the in vitro culture process [3]. The composition and concentration of growth regulators determines the direction of cell and tissue differentiation in vitro [4]. The cytoskeleton is a thin piece of tissue containing several layers of cells [5,11]. The thin cell layer is commonly cultured from leaves, stems, and inflorescences [6]. The thin cell layer usually contains many types of somatic cells, interspersed with cells capable of differentiating into shoot [7]. Using cell culture techniques to regenerate tissues and cells into complete plants for difficult-to-regenerate plants [8, 12]. There are least successful of Catleya sp. [13], Dendrobium sp. [14], Rhynchostylis sp. [15], Phalaenopsis sp. [16], Panax gingseng [17], Brassia napus [18], Petunia hybrida and Nicotina plumbaginifolia [19], morphogenesis [20], embrygenesis [21] and soot regeneration [22] in TCLs culture. This paper studies the technique of the thin cell layer culture on Dendrobium sp. and Cymbidium sp.

II. Materials and Methods

Study was carry out at Key Lab of Plant Cell Biotechnology (Institute of Tropical Biology, Vietnam Academy of Science and Technology) and Plant Biotechnology Lab (International University, Vietnam National University Ho Chi Minh City)