Optimizing the method of micropropagation of two native rootstocks of apple shortening (ornamental and jam)

Dissertation for receiving a master's degree in agricultural engineering

Biotechnology trend in agriculture

Abstract

This study was carried out in order to optimize the micropropagation method of two native apple rootstocks (ornamental and jam) and to improve their establishment and branching stage. First, samples were taken from healthy and active branches of the mother trees of these bases during the growing season. After preparing explants (side shoots and ends of healthy, young and mature branches) and their surface disinfection using sodium hypochlorite and Tween 20, nanosilver, they were placed on DKW, QL, WPM and MS media containing different amounts of cytokinins and auxins, in vitro conditions. After going through the establishment stage and choosing the best base culture medium, the micro-branches were used to optimize the steps of filling the branch and then rooting. In this research, the effect of different treatments on the establishment, processing and rooting of shoots, the effect of four basic culture mediums to obtain the best basic culture medium in the establishment phase of two varieties of jam and ornamental were investigated. In another experiment, the effect of three basic mediums QL, MS, WPM to obtain the best basic culture medium in the branching stage and also the use of different concentrations of BAP in combination with IBA in the branching stage were investigated. Also, two types of sugar sources, sucrose and sorbitol, the effect of different concentrations of IBA in combination with NAA on the rooting of several times planted explants (one-year explant) and explants after the establishment stage (new explant) were studied in Marbai variety. The results showed that in the establishment phase, the best growth parameters were obtained in the QL base culture medium. In the processing stage, the best growth type and the longest branch length were obtained for the explants cultivated in the WPM base culture medium. The highest number of branches was obtained in two culture media containing 0.5 mg/L BAP along with 0.1 or 0.01 mg/L IBA for the ornamental variety. The results of the present study showed that the highest number of leaves and the best growth appearance quality were obtained in the culture medium containing 30 grams per liter of sucrose compared to sorbitol for the Jam variety. Application of 4 mg/L IBA along with 0.25 mg/L NAA resulted in the highest percentage of rooting and root length for jam apple variety. The highest percentage of root formation, the highest number of roots produced in each explant, as well as the best index of apparent quality of explant growth were observed for explants grown several times (one year).

Key words: apple (Malus × domestica. Borkh), micropropagation, establishment, branching, rooting

Introduction

Humans have been trying to get food and shelter for thousands of years, and during this time they have made a lot of efforts to choose the best plants according to their needs, and fruits, vegetables and summer are also part of the food needs (Izdpanah, 2010). It is predicted that the world population will increase to 8 billion people in 2015 and to 10 billion people at the end of the first half of this century. If this forecast is fulfilled, the food production should increase twice the current amount in 2015 (Mirmohammadi Meibdi, 1382). Traditional techniques for plant breeding have made significant progress in breeding cultivars with high production potential, but these techniques are not able to increase the production of fruits and vegetables compared to the increasing demand for these products in developing countries. Therefore, there is an urgent need to use biotechnology to speed up the development of executive programs (Izdpanah, 2010).

Biotechnology is a new aspect of biology and agricultural science that provides new tools and solutions to solve the problems of food production in the world.The resulting new technologies were aimed at increasing the efficiency of production inputs (such as water, soil, manpower, energy, seeds and seedlings) and should be used in line with the use of minimal inputs with the highest production efficiency (Izadpanah, 2010).

In recent years, dense fruit tree plantations have been developed in the world, and instead of planting a few hundred trees per hectare, thousands of seedlings per hectare can be planted. cultivated In addition to that, recently home gardening [1] has also been noticed and developed in developed countries. To achieve this goal, the use of shortening bases in fruit trees has been considered in the past years. It is not possible to build uniform gardens using seed seedlings, so the production bases must be propagated and used by vegetative methods. In fact, the correct management of the garden increases the yield per unit area, and mechanized harvesting of the gardens. Finally, the use of resistant rootstocks in the garden in accordance with the climatic and environmental conditions will require the propagation of resistant, short and vegetative rootstocks (Izd Panah, 2010).

In order to mass produce the needed seedlings, the use of laboratory methods (tissue culture) has become widespread in the world, and several million seedlings and seedlings are produced annually in developed countries and some developing countries such as Turkey, Egypt, India, etc. In this regard, the propagation of shortened apple stems by tissue culture is necessary to produce uniform, cheap and short seedlings, and if you work on the bases used in home gardening, through the use of tissue culture techniques, you can achieve short and short pruning, which, while being ornamental and bearing good fruit, can also be used and cultivated in apartments (Kehrizi et al., 2016).

Theoretical basis of tissue culture technology It started with Gottlieb Haberland's idea about pluripotency (production of an organism from a single cell) in the early 20th century. Haberland stated that the techniques of isolation and culture of plant tissues should be developed that if the culture environment and nutrition of cultured cells are adjusted, these cells will repeat the stages of natural growth and development of plants in a few years. He predicted that tissue culture of plant embryonic cells can be successful. Therefore, he presented the concept of multiplicity. His advanced experiences before and after 1902 made Haberland justifiably known as the father of tissue culture. He thought that if all the necessary nutrients are provided to the cell, it can become a complete plant. The first successful tissue culture was done by White[2] in 1934. He reported the first successful carrot and tobacco callus culture until 1939. In fact, the advances after Haberland, who obtained more details about plant tissue culture, were achieved by White (1963), Rozdan[3] and Bojani (1983) (Kehrizi et al., 2016).

Micropropagation as one of the new methods of increasing the mass of plants, especially the bases of fruit trees, has been more attention in the last few decades. Production vegetative bases are often pollution-free, genetically pure and economically affordable, which are used to produce and build uniform and dense gardens. Since the basis of the construction of mechanized gardens is based on the production of vegetative and uniform seedlings, it is important to investigate suitable vegetative propagation methods, especially micropropagation, for the mass production of rootstocks (Dejmpour, 2016).

Vegetative propagation is one of the most widely used tissue culture technologies. To a large extent, the hereditary shedding of the plant is preserved.

2) The production of aberrant buds directly or indirectly.

3) Direct or indirect somatic embryogenesis, which has the potential to produce the largest number of plants, but is applicable in a small number of plant species.

Generally, in the conditions of in vitro culture [4] this possibility There is one that did the propagation operation throughout the year and permanently (Hartman[5] and Kester[6], 1990).

According to the available records and information, it can be stated that home gardening and the use of shortening bases have a significant contribution to the use of in vitro propagation methods and the mass production of healthy and uniform plants, with the help of which superior plant species in nature can be quickly propagated and from them. Used for correction programs or direct supply to the market.