| Populus Leucopyramidalis is a deciduous tree of populus euphratica and has unique value in wood production and urban greening.The breeding method of the seedling of the narrow-crowned poplar varieties is mainly used as a“pickling and picking”and“gun twist grafting”.The method is costly and long breeding period,which is difficult to meet the needs of the current large-scale cultivation,and its application is limited.In order to explore and establish a rapid breeding system of narrow crown poplar,and to promote the popularization and application of varieties.In this study,Agricultural University of Hebei cultivation of excellent poplar clones“HN-1”as the research object,the tissue culture technique was used to establish the rapid propagation system and leaf regeneration system,which laid the foundation for the rapid propagation and genetic transformation of the narrow poplar clones“HN-1”.The main findings are as follows:1.The young stem segments of the fine clones“HN-1”were used as explants to determine the appropriate basic medium and the best cytokinin concentration.Test results showed that,MS medium could be used as the basic medium for the proliferation of“HN-1”cluster buds,and the effect of cytokinin 6-BA concentration of 0.3 mg·L-1on the proliferation of cluster buds was significant.2.The effects of three plant auxin NAA,IBA,and IAA on the proliferation of poplar“HN-1”cluster buds were studied by orthogonal test design on the basis of screening out the concentration of cytokinin 6-BA.Screening results show,The medium suitable for“HN-1”cluster bud proliferation was MS+6-BA 0.3 mg·L-1+NAA 0.1mg·L-1+IBA 0.1 mg·L-1+IAA 0.15 mg·L-1.3.The two-factor test of NAA and IBA was set up with 1/2 MS as the basic medium.The results showed that,the suitable rooting medium was 1/2 MS+NAA 0.1 mg·L-1+IBA 1.0 mg·L-1.Simultaneous design of dark and normal light contrast test,proved that the conditions of the use of normal light rooting better,rooting rate of 71.1%.4.The experiment was carried out to carry out the experiment of seedling cultivation and field transplanting.The straw,vermiculite,loess and fine sand were selected as the transplanting medium.The results showed that the survival rate of transplanting seedlings was 93.8%,and the survival rate of the transplanted seedlings was as follows:peat:loess:fine sand=1:1:1（volume ratio）.5.The leaves of“HN-1”were used as explants to establish the regeneration system of leaf genetic transformation.The suitable medium for differentiation of adventitious buds was MS+6BA 1.0 mg·L-1+NAA 0.1 mg·L-1,and the germination rate was89.33%.The effect of 50 mg·L-1 kanamycin（Km）on leaf differentiation was the most significant.The addition of 300 mg·L-1 cefotaxime sodium（CTX）in the culture medium could effectively inhibit the growth of Agrobacterium and promote the differentiation.6.Based on the establishment of the“HN-1”tissue culture rapid propagation system,combined with the market price of each reagents,the cost of the“HN-1”tissue culture seedlings was put into the factory.Annual production of 200,000 narrow-crowned poplar“HN-1”seedlings of tissue culture plant nursery costs:The cost of plant tissue culture seedlings was 0.4942 yuan per plant.The cost of transplanting the bottle seedlings per bottle is 0.2610 yuan per plant.The cost of indoor infrastructure and equipment depreciation of the plant tissue culture plant and the damage of the culture vessel were0.0930 yuan per plant.The outdoor part of the plant tissue culture plant and equipment depreciation,heating and other costs of 0.0500 yuan per plant.The final cost per plant tissue culture seedlings is 0.4942 yuan per plant+0.2610 yuan per plant+0.0930 yuan per plant+0.0500 yuan per plant=0.8982 yuan per plant. |
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