Optimization of Factors Affecting in vitro Propagation of  Myrobalan 29c Rootstock

This study was conducted aimed to consider the effects of culture medium and the concentration of growth regulators on proliferation, rooting and the acclimatization of Myrobalan29C. This study was performed as a factorial experiment in a completely randomized design (CRD) with four replications where each plot contained five explants. Given the role of trees rootstock of growth rate vegetative in the early maturity٫ yield and disease resistance will be suitable rootstock that has an important role in program garden management. In total rootstock of fruit trees have be propagation sexual and asexual methods, however٫ given that in sexual reproduction the resulting dispersion characteristics and the resulting seedlings changed by genetics, it is tried for decades to asexual propagation methods in specially tissue culture methods healthy rootstock for mass propagation and used the development of orchards. Seedling rootstock Myrobalan had been used in the past. The rootstock can be positive features cited the ease of access, to be cheap, good yields after maturity. In study, the effects of NAA and BAP on proliferation of Gisala5 rootstock showed the most shoot treated in mediacontaining 1mg/l BAP. Investigated Chinese plum in vitro micropropagation showed that 1/2 MS media had the highest percentage rooting, and acclimatization rooted plantlets to greenhouse conditions was using the system miss successfully. Due to the importance and essential to achieve an efficient protocol for the mass propagation of Myrobalan 29C in Iran, this study was conducted with the main purpose of evaluating the most suitable media culture and plant growth regulators in micropropagation of Myrobalan 29C.
Material and The explants were collected from shoots of Myrobalan 29C rootstock maintained in the experimental greenhouse of Khorasan Razavi Agricultural and Natural Resources Research Centre (of Mashhad, Iran), on June 25, 2013. The explants were washed by water and dishwashing liquid to removed surface contamination. Then they were divided to some parts containing one bud and were sterilized with ethanol 70% 1 min and sodium hypochlorite 10% at 10 min. Proliferation was performed in two kind of culture medium (MS and DKW) that supplemented with plant growth regulators BAP (0, 1, 2, 3, 4 mg l-1). The rootstocks of in this step, after subculture three (21 days between each subculture), the numbers, the length and quality of the shoots (explants strong growth, with no signs of vitrification, necrosis of leaf are yellowing terminal meristem), b– less than 15% have the symptoms of vitrification, necrosis of leaf are yellowing terminal meristem, and c– explant weak, 15-30% have the symptoms of vitrification, necrosis of leaf are yellowing Terminal meristem) were measured. This stage was carried with four replications and each replicates with five samples. Two culture media (MS and DKW) were used for rooting, which supplemented with indole-3-butyric acid (IBA) at four levels (0, 1, 2, 3 mg l-1). This stage was carried with four replications and each replicates with three samples. After being rooted explants, the best cultured media and combination of rooting growth regulators number and root length, leaf number and stem length and quality of explants were recorded. Acclimatization used in substrate including Coco Peat - Perlite 3:0/5 V, Coco Peat 100%V٫ Coco Peat - Perlite 0. 5:3V, Perlite100%V and Coco Peat - Perlite1:1V).
Results showed that the 10% sodium hypochlorite for 10 min, with 4% decay was the best treatment for sterilization. The results showed that the proliferation average was 6/16 in MS medium with 2 mgl-1 BAP and the most percent of rooting and root length were about 100% and 2/51 cm in MS medium with 2mgl-1 of IBA, respectively. The acclimatization of plantlets to greenhouse conditions was successful. The highest rate of plantlets survival (about 70%) was obtained from substrate Cocopit and Prlit (1; 1 V). In the present study, explants year compared annual explant least contamination had enjoyed. It seems that the young explants, smooth the surface of the skin, having the least amount of crack depend on the type of surface depressions explants and the crack and lower depressions the surface explants increase surface contact area disinfectants and also improve its impact. In this study found that the type of medium a significant impacted on the health of plants and so the proliferation of explants was successful. Usually root production in plants under the influence of synthesis, metabolism, and transport is auxin signaling pathways. Therefore acclimatization directly affected the rooting of plants that had high quality and the best rate of induction.
The results of this research showed that we can duplicate Myrobalan29c rootstock by in vitro method. According in this research, MS media including BAP and IBA plant growth regulators are the most suitable for micro propagation.