Study on Technology of Immature Culture and Callus Inducing from Young Segment of Pinus Massoniana
|School||Central South University of Forestry Science and Technology|
|Keywords||Mason pine Immature embryo Young stem segments Organogenesis Plant regeneration Callus culture|
Masson pine (Pinus massoniana Lamb.) Is a the important local fast-growing conifers, higher utilization value, explore the trees in vitro propagation techniques, the establishment of a rapid propagation system, genetic improvement of this species, and research in genetic engineering good work laid the theoretical and technical basis. Immature embryos of masson pine seedlings tender stem segments as explants, organogenesis and callus culture studies to explore the impact of different factors on the organ or callus. The main findings are as follows:. The Masson pine embryo and young stem segments as explants, the sterilization process, were they different processing and comparative analysis of their respective off bacterial effect, the results showed that: in the the two explants different processing, alcohol sterilization process compare significantly, sterilization time is too short, incomplete sterilization, explants susceptible to infection; sterilization time is too long , although the good sterilizing effect, but because explants ter susceptible to damage, resulting in death of explants. Masson pine immature embryos sterilization process, the best method of disinfection: 75% alcohol surface sterilization 30s, then 0.1% mercuric chloride 8min after the last rinse 5-6 times with sterile water; of cauda equina The pine seedlings sterilization best method is to use 75% of the alcohol-soaked 20s, soaked in 0.1% mercuric chloride 8min, final rinse 5-6 times with sterile water, the pollution and mortality rates are the lowest, only 3.71%. Pinus direct organogenesis culture prevention of browning effective way is added to the medium 0.15g/LVc, and two days Adapter to shorten the residence time of the explants in the original medium , reduce browning chance. At the same time through a lot of comparative analysis, draw: the initial culture phase the WPM medium of germination rate in medium GD and DCR, and better growth conditions. BA, KT and NAA most suitable concentration level of 1 mg / L the 0.5mgm and 0.15mg / L NAA excessive callus degree increase. Concrete more suitable for the medium induced by a combination of Dendrolimus shoots WPM lmg / L BA 0.5mg / L KT 0.15mg / L NAA 0.15g / L Vc; proliferation and elongation of multiple shoots culture, the most suitable medium combination WPM 0.15mg / L NAA 0.15g / L Vc 0.5g / L Ac; rooting culture, the best medium combination WPM 2mg / L IBA 0.15mg / L NAA 0.5g / L Ac add rooting medium sucrose concentration to 20g / L appropriate. 3. In Masson pine callus culture process, the ratio of cytokinin and auxin explant dedifferentiation of callus formation, plays an important role in regulating. Therefore must constantly adjust the proportion between the various hormone concentrations, in order to facilitate the Masson pine callus induction, the test derived induced Masson pine callus medium combination GD 2.5mg/LNAA. The same time, through a lot of trials and research analysis found that of NAA plays a major role in Masson pine callus induction and lack of NAA, callus is difficult to form, which can be seen, NAA induced callus The essential factor. Through a lot of comparative analysis, we have come to Masson pine immature embryos of masson pine than tender stem segment regeneration ability, and relatively easy to induce differentiation of the regeneration plantlets.