Determination of an effective method of Citrus aurantifolia lime and Citrus lemon varieties plant explants sterilization for in vitro introduction into the culture

Abstract

Potted citrus plants are in constant demand, but the propagation of such plants requires the use of a winter greenhouse and several well-developed mother plants to obtain planting material. The use of microclonal propagation can accelerate the production of seed material though it requires detailed development of in vitro cultivation techniques. The aim of the research was to study the effectiveness of sterilizing substances and methods of sterilization on the yield of viable micro shoots of lime plants (Citrus aurantifolia) as well as Meyer and Jubilee lemon varieties and their growth in vitro. The research was conducted in the interdepartmental "Biotechnological Laboratory" of the Agrobiotechnological Faculty of Bila Tserkva NAU. The source material was microshoots from lime plants (Citrus aurantifolia), and lemon varieties (Citrus lemon) Meyer and Jubilee. The objects of the study were selected according to the diversity of genotypes, development type and varietal characteristics. The possibility of using different parts of plants for reproduction is a feature of in vitro culture. We used microshoots with a bud in our research. To neutralize the exogenous bacterial and fungal microflora, we used 70 % ethanol solution C2 H5 OH, sodium hypochlorite 5 %, 15 % solution of hydrogen peroxide H2 O2 , sulema (0.1) and washed the material in sterile water (for 5–10 min) under several schemes. The study revealed the influence of different sterilization regimes for obtaining sterile explants and the introduction into plant culture of explants of the species of lime Citrus aurantifolia as well as Meyer and Jubilee lemon varieties. It was found that the use of step sterilization using 2.5 % solutions of sodium hypochlorite gives a high percentage of sterile viable material – 27.7 %, which is significantly higher than in other variants of the experiment. Key words: microshoots, lime, sterile material, micropropagation, sterilization, hydrogen peroxide, viability of explants.