| Almond(Amygdalus communis L.)is a temperate stone fruit of great value in food industry, medicine and medical treatment, which is conventionally propagated by grafting. The micropropagation procedures of two kinds of Almond's rootstocks Nemaguard and Lovell had been researched, including the establishment of aseptic cultures, shoot proliferation, selection of suitable medium and hormone, rooting and acclimatization. A feasible and practicable micropagation process of these two rootstocks had been made. High survival rate of the explants can be achieved when Nemaguard and Lovell's explants were collected in the April and June. Washing the explants under running tap water to remove some of the dirt and contaminations. The stems,with single bud,were cultured on Murashige and Skoog(1962) medium (MS) added with 0.5mg/L N6- benzyladenine (6-BA) and 0.2mg/L gibberellic acid (GA3) for Nemaguard,MS added with 2.0mg/L 6-BA and 0.1mg/L GA3 ,the survival rate were 73.3% and 80.0%.During the shoot proliferation,we found that MS basal medium was more suitable for growth and subculture of the shoots than MS-R,Gs and H basal mediums.The exogenous application of cytokinins were very necessary for shoots proliferation,the multiple effect of 6-BA was better than kinetin(KT). Combinations of cytokinin and auxin can stimulate the growth of shoots,and the effect of indile-3-acetic acid (IAA) was better than indole-3-butyric acid (IBA) and naphthalane acetic acid (NAA).0.1mg/L IAA can improve the proliferation rate for Lovell,0.1mg/L GA3 improved the proliferation and made the shoots growth better for Nemaguard.100mg/L lactalbumin hydrolysate (LH) improved the proliferation and made the shoots growth better for two rootstocks.For Nemaguard,the optimum proliferation medium was MS basal medium supplemented with 0.5mg/L 6-BA,0.1mg/L GA3 and 100mg/L LH; and MS basal medium supplemented with 1.0mg/L 6-BA,0.1mg/L IAA and 100mg/L LH was also found to be favorable for Lovell's proliferation.It was good to keep the culture at about 27℃ on a 24hr photoperiod with a light intensity of 2,500 Lx on the cultures,and to subculture Lovell's cultures every 30 days,Nemaguard's cultures every 40 days.The proliferation rate were 3.83 and 3.05.MS basal medium added 0.3mg/L 6-BA,0.1mg/L GA3 and 0.1mg/L IAA made the shoots elongation and better growth.One-half MS basal medium was more favorable on root formation than MS medium for rooting.The rooting rate was increased significantly when we added auxin in the medium.It was good to use 0.5mg/L IBA to induce root,and cultured in dark 10 days,The rooting rate of Nemaguard and Lovell were 80.0% and 68.2%.0.1% activated chaicoal (AC) improved to root.Socking of the base od the shoots in 100mg/L IBA solution for 60min,then transplanting into 1/2MS medium without hormones obtained over 50% rooting rate.Three-steps acclimatization procedure was used to acclimatize the plantets successfully to free living condition. The survival rate were over 80%.The histological and morphogical study of orgin and development of adventious root of Almond's rootstocks plantet in test-tube was conducted.The histological section observation in each period of adventitious root origin was the same in the two varieties. It was connecting place of cambium and pith ray that the adventitious root primordium was formed. The adventitious root primordium differentiated on the 6th days.Later the tissue differentiation of the roots gradually appeared.The vascular tissue of the root and the stem joined quit well.SDS polyacryla mide gel eletrophoresis was employed to analyse the soluble protein zones of leaves of two almond's rootstocks species.The result shows that there are differences in constituents of protein zons among different wild tree peony species.Two almond's rootstocks species the band which exhibited unchangeable more or less from beginning to the 12th culture.It shows that tissue culture keeped specific genitic stabilization,was a good method of propagation. |
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