| In order to accelerate the speed of propagation of Lilium rosthornii and shorten its reproductive cycle, and thus for the purposes of maintaining the excellent germplasm and traits of Lilium rosthornii, and so as to protect the biodiversity of the Liliaceae, in this study, scales, leaves, stem segments, flower organs, renewable leaves and bulblet were used as explants, which were inoculated in cultured medium supplemented with different concentration and composition, such as6-BA and NAA. This research studied the major factors influencing the tissue culture and rapid propagation of Lilium rosthornii, and mainly explored the best explants, medium components and culturing methods, so as to establish a complete regeneration system of tissue culturing of Lilium rosthornii. The results showed that:l.With the scales, leaves, stem segments, flower organs, renewable leaves and bulblet as the explants, the results were as follows:the higher contamination rate of scales, leaves and stems, the lower callus formation rate would be; the ovary and receptacle of floral organs had meristematic capacity, with a high rate of induced differentiation and more budding, the induced effects of receptacle was better than the ovary, but other parts of floral organs had a high rate of mortality; Renewable leaves and bulblet of aseptic seedlings were not contaminated, with a large number of base and higher proliferation rate. So the receptacle, renewable leaf and bulblet were selected as the best induced explants.2.In primary culture, the best medium for induced cluster seedlings of scales was MS+6-BA1.5mg/L+NAA0.5mg/L(the sugar was30g/L and the agar was7g/L), with a differentiation rate of82.33%and differentiation coefficient of3.33; With stems as explants, the best medium for cluster seedlings was MS+6-BA1.5mg/L+NAA1.0mg/L, with a differentiation rate of58.67%and differentiation coefficient of3.19; With ovary as explants, the best medium for cluster seedlings was MS+6-BA1.5mg/L+NAA0.2mg/L, with a differentiation rate of93.33%and differentiation coefficient of3.4; With the leaves of regeneration plants as explants, the best medium was MS+6-BA1.0mg/L+NAA0.2mg/L, then the differentiation rate and differentiation coefficient were54.33%and2.9; With the bulblet of regeneration plants as explants, the best medium was MS+6-BA1.0mg/L+NAA0.5mg/L, then the differentiation rate and differentiation coefficient were100.00%and2.5.3. On the proliferation cultured stage, cytokinin played a significant role on multiplication coefficient. The appropriate medium for proliferation of adventitious buds was MS+6-BA1.0mg/L+NAA0.05mg/L, where the buds grew well and the multiplication coefficient was3.1. There were more callus, which had great potential of continuing to divide the adventitious buds. The height of formation plant was uniform, bulbs also enlarged significantly.4. Low concentrations of inorganic ions were good for the rooting of Lilium rosthornii, so this study selected1/2MS medium. NAA was better than IAA in promoting root induction, the best concentration was0.5mg/L; the best concentration of sugar for rooting was40g/L; the best concentration of activated charcoal for rooting was1.0g/L; PP333(10.0mg/L) can dwarf and strong seedling, now rooting rate was86.67%, average root number was5.2, the plant had developed root system and deep green fleshy leaves, which provided a guarantee for later transplanting seedling. Considering various factors, the best rooting medium was1/2MS+NAA0.5mg/L+10.0mg/LPP333+40g/L (sugar)5. The concentration of sugar also affected the expansion of bulbs. The results of this study showed that:the concentration of30-80g/L sugar had the fairly good effect on enlargement of bulb. Within this range, the diameter and weight of young bulbs were enhanced with the increase of sugar concentration. Higher concentration of sugar (more than80g/L) in medium would inhibit seeding growth. So80g/L sugar concentration was more conducive to the enlargement of Lilium rosthornii bulbs. |
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