Somatic Embryogenesis And Cell Suspension Culture Systems Of Juglans Mandshurica

Juglans mandshurica Maxim. subordinates to juglandaceae is Deciduous trees, originated in the Yellow River north and northeast. J. mandshurica Maxim is dubbed one of the "northeast three hardwood" (Fraxinus mandshurica Rupr., J. mandshurica Maxim., Phellodendron amurence Rupr.). Due to the strong branches and big leaves have the ornamental value and with good resistance and patience, also can absorb the industrial waste gas to improve the human living environment. Juglans mandshurica is broadly used in application of landscape recently, mainly street greening and small environmental group of plant. Besides J. mandshurica Maxim. also has high pharmaceutical and industrial production value. Due to the excessive J. mandshurica Maxim. resources exploitation and seed bank cause low reproduction rate of the lack of J. mandshurica Maxim. forest resources. Somatic embryogenesis training for the conservation and utilization germplasm resource provides a new way. This thesis mainly discusses the occurrence of somatic embryogenesis and cell suspension culture system of J. mandshurica Maxim. In order to realize the rapid propagation, thoroughbred cultivation, somatic embryogenesis bioreactor production technology and forest tree genetic engineering research laid the foundations, the main research results are as follows:1. The best condition of induction J. mandshurica Maxim. embryo callus is:①zygotic embryos for explant is most easily to form sexual embryo callus, the best time of gather materials for explant is in May to June, Zygotic embryos callus induction rate topped 91.3%.②the optimal nutrient medium for induce callus is MS+1.0mg·L-1 2,4-D+0.5mg·L-1 6-BA with 30g/L sucrose and 6g/L agar.③somatic embryogenesis induction, development and differentiation of appropriate medium is MS culture additional sucrose 60 g·L-1,700mg·L 1casein acids hydrolysate,6g/L agar,without any growth regulating substances.2. Embryo callus turns to 1/2MS liquid medium adding 0.05 mg·L-12,4-D.1/4MS medium without any growth regulating substances is conducive to the development of somatic embryos and maturity. In liquid cultivate process,7g/L vaccination density promoted the formation of somatic embryogenesis,3. In the cells suspended medium of J. mandshurica Maxim., pH change trend in a "V" word, cell dry weight in "S" value change shape curve:after inoculation first 6d, the pH average dropped to 4.21, cell dry weight slow growth; In the 12th d the pH averages gradually rose to 5.3, cells grown rapidly dry weight; After 15d cultured in the suspension cell growth end, the PH average maintain at 5.44 around, cell dry weight maintain stably.4. In cell suspension culture process:①In MS liquid suspended medium adding 3% sucrose is good for cell growth, sugar concentration too high or low influence the cells to grow.②The suspension cultured cells in about 15d growth cycle.③Under different sugar concentration suspension culture found that the speed of sugar hydrolysis only in adding time growth with sugar concentration.④the content inside cells with the reducing sugar changed with the sucrose content, the otherness decrease gradually.