The Research of Tissue Culture in Fast Breeding Technology for Osmanthus Fragrans Lour
|School||Central South University of Forestry Science and Technology|
|Course||Tree Genetics and Breeding|
|Keywords||Osmanthus Fragrans Lour stem embryo leaf tissue culture|
The experiment purpose is to do the research of Osmanthus fragrans Lour. plant tissue culture which happened in variety of ways. The experiment contained two facets of tissue culture research.One facet is that taking the young tress,germfree seedling stem and embryo as explants to regenerate in the form of germfree short tress cutting.The other is that taking young leaf,embryo as explants to occur in the form of callus.The results are as follows:1 Tissue culture research of regenerating in the form of germfree short tress cutting(1)Germfree system established The seed which inoculated at low contaminat-ion rate is easy to disinfect. Using Hgcl20.1% to disinfect young new tress for 3-6 minutes is available.The top bud is at low contamination rate.(2)Initial culture comparing with medium B5 and WPM, medium MS is more available for embryo initial germinating;Less than the dosage of 1×1014 N+·cm-2 is available for embryo germinating when N+ immitting in the embryo body.Dealing with the seed in the environment of low temperature 4℃and cold storage for 210 days can break embryo dormancy in effect,accelerating the embryo to bourgeon in advance. Comparing with medium B5 and MS, medium WPM is more available for young stem germinating;While controlling the stem not to be browned,the AC concentration should be controled within 0.1%.(3) Multiplication culture for embryo plants B5+0.50 mg·L-1TDZ+0.05 mg·L-1 NAA is available for tuft buds inducement. B5+5.00 mg·L-1VBA+0.10 mg·L-1NAA, B5+1.00 mg·L-16-BA+1.00 mg·L-1GA and WPM+0.50 mg·L-1 TDZ+0.05 mg·L-1 NAA+0.50 mg·L-1BR are available for elongation and multiplication culture of embryo plants.The plant with ion beam implantation in embryo is at about 2.04 cm high in elongating culture process; The plant with no ion beam implantation in embryo is at about 2.77 cm high in elongating culture process; The diversity between each other is notable(4) Multiplication culture for stem plants B5+7.00 mg·-16-BA+0.10 mg·L-1 NAA,B5+0.10 mg·L-1TDZ+0.05 mg·L-1NAA are available for tuft buds inducement. B5+8.00 mg·L-1KT+0.05 mg·L-1NAA,B5+2.00 mg·L-16-BA+1.00 mg·L-1GA,B5+2.00 mg·L-16-BA+0.10 mg·L-1BR,WPM+0.50 mg·L-1TDZ+0.05 mg·L-1NAA+0.50 mg·L-1 BR and B5+2.00 mg·L-6-BA+3.00 mg·L-1Biotin are available for stem elongation inducement.(5)Rooting culture 1/2MS+0.60 mg·L-16-BA+4.00 mg·L-1 NAA is available for rooting culture inducement.2 Tissue culture research of occuring in the form of callus(1)Initial callus inducement culture WPM+0.12 mg·L-12,4-D is available for embryo callus inducement. B5+0.12 mg·L-12,4-D+1.50 mg·L-16-BA is available for young leaf callus inducement. B5+5.00 mg·L-16-BA+0.10 mg·L-1NAA is available for young sterile plant leaf callus inducement.(2)Multiplication callus inducement culture WPM+0.50 mg·L-1TDZ is available for embryo callus multiplication. WPM+0.10 mg·L-1BR+0.10 mg·L-1 NAA is available for leaf and embryo callus multiplication.(3) Polarization callus inducement culture B5+0.40 mg·L-1TDZ+0.10 mg·L-1 +0.50mg·L-1BR is available for embryo callus polarization. WPM+1.00 mg·L-1TDZ +0.05 mg·L-1NAA and B5+5.00 mg·L-16-BA+0.10 mg·L-1NAA are available for leaf callus polarization.(4)Rooting culture 1/2MS+0.60 mg·L-16-BA+4.00 mg·L-1NAA is available for rooting culture inducement.