| Vaccinium Uliginosum is one of the most important wild resources in China,with a variety of specific quality,which is an important resource for cultivating new varieties.But there is no artificial cultivation,artificial propagation under natural conditions are very difficult,making exploitation of this valuable natural resources are extremely limited.Plant tissue culture can solve the puzzle,but at home and abroad on other blueberry tissue culture research has become more mature.The cultivation of Vaccinium Uliginosum and the plant regeneration and so far is rare.In this experiment,using the wild Vaccinium Uliginosum for the test materials,the effects of plant growth regulators,dark culture time,p H and light on the callus induction of the leaves and the buds were studied by using the tissue culture method.And the effect of different hormones ratio and different hormones on the direct regeneration of adventitious buds in leaves and non-bud segments callus and on the regenerated plant taking roots were studied.The aim was to establish a complete system of regeneration of Vaccinium Uliginosum,And lay the theoretical basis to the next step in tissue culture breeding and genetic engineering breeding.The experiment results are as follows:(1)The optimum callus induction medium was: WPM + 3 mg / L ZT + 0.2 mg / L NAA + 20 g / L sucrose +8 g / L agar.The callus induction rate of leaf and stem was up to 100%.The callus color was fresh green and the texture was compact.The callus of stem was better than that of leaf.The optimum induction p H of leaves was 5.2.The optimum illumination time was 10 hours per day.The best dark culture time was 10 days,and the callus induction results in different positions was: petiole> leaf> whole leaf cutting blade> tip,And the callus induction effect of blade tails placed was better.The optimum induction p H of stem segment was 4.8,the best illumination time was 12 hours per day,and the best dark culture time was 20 days.(2)The best adventitious bud induction medium of Vaccinium Uliginosum was: WPM + 3.0 mg / LTDZ + 0.5 mg / L ZT + 0.1 mg / L IBA + 0 mg / L NAA + 20 g / L sucrose +8 g / L agar.The The differentiation rate of leaf callus was up to 100% and the differentiation rate of stem callus was 93%.(3)Effects of different components of ZT,IBA and sucrose on callus differentiation: The optimum medium for adventitious bud differentiation of leaf callus was: WPM + 3.0 mg / L ZT + 0.01 mg / L + 25 g / L sucrose +8 G / L agar.The optimum adventitious bud induction medium for stem callus was WPM + 2.0 mg / L ZT + 0.01 mg / L + 25 g / L sucrose +8 g / L agar.The callus differentiation time was longer in this condition,and the best induction rate of leaf callus was higher than that of stem.The high concentration of sucrose inhibited the differentiation of stem callus.(4)When the cytokinin was used alone,the induction effect was: ZT > TDZ > 6-BA,and the effect was not as effective as combination groups.ZT is suitable for adventitious induction.The induction time quite short,only two weeks can induce adventitious buds,and adventitious buds can grow well;TDZ induced more buds,but TDZ inhibit bud elongation growth;6-BA is not suitable for the induction of adventitious buds and the induction effect is poor.(5)The optimum rooting induction medium was1 / 2 WPM + 1.0 mg / L + 0.1 mg / LNAA + 30 g / L sucrose + 8 g / L agar,and the rooting rate was 84%.When IBA and NAA were used alone,the rooting effect of IBA was better than that of NAA,and the induction rate of IBA 5.0 mg / L was 87% higher than that of combination medium,and the root and seedling growth were the best. |
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