| Koyama spruce was fine tree species in planting timber forests,planting protective forests and greening courtyard,with beautiful appearance,fine texture,straight stem,fast growth and high adaptiveness,has high economic value,ecological value and aesthetic value.Forest areas of northeast china are promoting energetically the development of koyama spruce plantations in recent years.The total area of koyama spruce plantations covers about 5%of the total plantations area in daxinganling forest region and will be rised up to 20%after ten years,will exceed the area of Larch plantations.The little block of koyama spruce natural forest shows punctate distribution and this situation is tending to shrink.Some high quality germplasm resources of koyama spruce have been lossed seriously.The high quality seed orchards have not formatted productivity.Scope of the selection and forestation f the superior clone of koyama spruce is relatively small.The many seedlings have not been progeny tested from natural forest and plantations to be used at present.The protection,preservation and breeding of high quality germplasm resource is relatively lagging.The research level of koyama spruce falls behind western developed countries greatly at the same kind tree species,such as picea abies,picea glauca and picea mariana.Therefore,the study on somatic Embryogenesis and formation of adventitious buds in Picea koraiensi is very exigent.This study not only may solve the problem of excellent seedings breeding and germplasm resource preservation but also lay the foundation for the research of the genetic engineering technology in koyama spruce.The main results are as follows:The relatively good embryogenic callus induction have been achieved on the improved basic medium RJW supplemented with 0.46mg·L-1BA,0.53 mg·L-1KT,3.07 mg·L-1NAA,20g·L-1 sucrose, 450mg·L-1 L- glutamine,750mg·L-1 casein hydrolysate,2 g·L-1 gelrite under the 22℃,dark culture condition from immature zygotic embryos of koyama spruce.The average induction frequency was 81.27%.The genetic basis of koyama spruce has significant influence on embryogenic callus induction.The highest proliferation rate and proliferation quantity of koyama spruce embryogenic callus have been achieved according to the mode of solid and filter paper alternate culture on the improved basic medium RJW supplemented with 0.21mg·L-1BA,0.20 mg·L-1KT,1.85 mg·L-1NAA, 5g·L-1 inositol,20g·L-1 sucrose,450mg·L-1 L- glutamine,750mg·L-1 casein hydrolysate,2 g·L-1 gelrite under the 22℃,dark cultivation condition.The relation among induction rate,proliferation rate and proliferation quantity of embryogenic callus was significant positive correlation.The proliferation frequency and proliferation quantity drop gradually along with the increase of subcultured times.The different cell lines have different descending rate.The best effect of globular embryos induction achieved on the improved basic medium RJW supplemented withl 5g·L-1 inositol, 3g·L-1 activated carbon and without any plant growth regulators.The average formation rate was 97.6 entries/g.The response of three cell lines with different genetic basis was uniform to middle concentration of inositol and activated carbon.The relatively good somatic embryos maturation achieved on the improved basic medium RJW supplemented with 7.5mg·L-1ABA,40g·L-1 sucrose, 20g·L-1PEG4000,1.76 mg·L-1 vitamin c.The somatic embryo maturation rate of 3#,2# and 9# was up to 45,65 and 15 entries/g,respectively.The average rate was 41.7 entries/g.AgNO3 not only did not promote the differentiation of koyama spruce somatic embryos but also played inhibitory effects to the differentiation.The highest percentage of somatic embryos germination achieved on the improved basic medium 1/2RJW supplemented with 3g·L-1 activated carbon.The percentage of somatic embryos germination was 88.5%.The optimum basic media was 1/2BMI in conversion stage to somatic embryos desiccated for eight days.The average conversion rate was 18%.The highest adventitious bud callus induction rate achieved on the basic medium SH supplemented with 0.2 mg·L-1BA,0.4 mg·L-12,4-D under the 22℃,light culture condition from immature zygotic embryos of koyama spruce collected at august 3.The adventitious bud differentiation and adventitious bud quantity were all the best on the basic medium SH supplemented with 0.2 mg·L-1BA,0.2 mg·L-12,4-D.The adventitious bud differentiation frequency and adventitious bud quantity was up to 90.2%and 15.4 entries,respectively.The only basic medium 1/2BMI was the best on adventitious bud elongation.The adventitious bud elongation rate was up to 57.9%.The seedling height growth was up to1-1.5cm during 3-5w.The basic medium 1/2BMI without any plant growth regulators was optimum to rooting induction.The rooting rate was up to 14.6%.The genetic basis has a great effect on rooting of seedling.3# has the highest rooting rate for 34.7%.
|
PDF Full Text Request