| Hazelnut as one of the world's four major nuts, it had the advantages of unique flavor, nutrient-rich, high economic value. The hybrid hazelnuts had characteristic of domestic hazel like cold hardiness and suckering strongand also had advantages of european hazelnut like nuts big and the high yield. So they had been widely promoted. Currently the reproductive system of hazel was cuttage propagation and mound layering etc. Due to its low propagation coefficient, it unable to meet the market demand for hazelnut seedlings increasingly. Tissue culture was a breeding technology of high efficiency and low cost. Using to hybrid hazel (84-349,84-226,84-69) as the tested material. On the one hand, to do further tentative research and optimization for the tissue culture of subculture and rooting, further more to propose the bottle rooting, on the other hand, to make a preliminary study on callus induction and differentiation for hazel stems, leaves and cotyledons and it laid the foundation for hazelnut tissue culture regeneration system. The main results are as follows:(1) Improved NRM medium as a subculture medium was more suitable for the growth of84-226.Choose6-BA2.5'mg·L-1and IBA0.01mg·L-1as the subculture hormones. The older generation to reduce its concentration to6-BA2.0mg·L-1and IBA0.005mg·L-1that help to promote the proliferation of hazelnut stems.(2) TDZ appropriate concentration range was0.001mg·L-1-0.01mg·L-1and0.005mg·L-1was the best.GA3was the role of promote the sprouts elongation, but the proliferation of GA3on the hazel stem segments had no significant effect. The best experimental concentration was0.1mg·L-1. When the concentration reach0.3mg·L-1, the growth of sprouts was inhibited. Polyamines have significantly effect higher than GA3, but both added was not better than a single addition of polyamines on the proliferation of the stem segments.(3) With the following the Generations increase, the continued use of IBA and6-BA would cause the increase of IAA,6-BA also contributed to formed callus of the stem segments on roots and appeared buds. Also it caused changes in the sprouts body of the ABA. In the overall, GA and ZR had smaller changes and it had no obvious with low levels on proliferation of stem segments.(4) The rooting effect with NAA was better than IBA and no significant difference between add the two and separate the addition of NAA. For consideration of production, the best rooting treatment was1/2DKW+NAA0.2mg·L-1. Non-tube rootage with high rooting rate, short breeding cycle, saving production costs and other advantages, but it was not perfect about management program on hardening transplanting.(5) As the different month of endogenous hormones in bottle rooting of tissue culture and PPO,POD activity were shown a some changes, and it showed consistency with survey conditions of greenhouse transplanting growth(6) The NRM+2,4-D1.0mg·L-1+IBA0.5mg·L-1was the best callus induction medium for hazelnut stem and leaf, and add PVP antioxidant effect than the VC and AC. Callus induction stage, the hormone combination induced effect:auxin and auxin combination>cytokinin and cytokinin combination>auxin and cytokinin combinations.(7) MS+2,4-D4.0mg·L-1+6-BA0.5mg·L/-1+Proline500mg-L-1was the best callus induction medium for hazelnut cotyledons, hormone of callus induction effect which combination with2,4-D is6-BA>TDZ>NAA>KT.(8) Stereoscope observation the form built of callus induction and differentiation. Basic line was postembryonic development. We observed its growth morphogenesis by scanning electron microscope. |
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