Study On Technique Of Tissue Culture And Micropropagation Of Pinus Bangeana And Pinus Ponderosa

The study on the tissue culture and micropropagation of Pinus bangeana and Pinus ponderosa showed that: under vitro condition,the mature embryo of Pinus bangeana was introduced in large quantity of adventitious buds, the reproduced plantlet was regenerated from the mature embryo of Pinus ponderosa with great amount of rooting rate. The effective way for micropropagation of Pinus Bangeana and Pinus ponderosa was tentatively explored. The main results are as the following:(1) The embryo is an optimum explant for micropropagation of Pinus bangeana and Pinus ponderosa. The explant growing and adventitious buds differentiation were well when the embryo root was cut before inoculation.(2) Pinus bangeana seeds were soaked in hot water with 55℃ for 30min, and then soaked in normal water for 3 to 5 days; the seeds of Pinus ponderosa were soaked in hot water with 55℃ for 10min, and then soaked in water for 2 to 3 days; The well-marinated seeds were sterilized by using 0.2% KMnO4 to Pinus bangeana for 30min and to Pinus ponderosa for 10min, then seedcoat was removed. The embryo was soaked in 70% alcohol for 20s, then transferred into 0.1% HgCl2 for 8 to 10min, and washed by sterile water for 4 times; under disinfective condition, the endosperm was striped by using little dissection knife to get mature embryo into adventitious buds medium. The embryo is rarely polluted.(3) MS is the best adventitious buds inducement medium for Pinus bangeana. The adventitious inductive rates reached to 70%, it is also used into adventitious buds propagation and growing.(4) GD is a best medium for adventitious buds inducement to Pinus ponderosa, 1/2GD and 1/2SH were a better medium for adventitious propagation, GD and SH were better medium for adventitious growing, 1/2 GD was a better medium for roots inducement among four mediums.(5) The inductive rate of adventitious buds for Pinus bangeana reaches 70% above by using 6-BA 3.5 mg· L-1combing NAA 0.4 mg· L-1 with IBA 0.1 mg· L-1. (6) The inductive rate of adventitious buds for Pinus ponderosa reaches 60% above by using 6-BA combing NAA with lower hormone level at 6-BA 0.5—1.0mg·L-1 and NAA 0—0.5mg·L-1; Which could accelerate the adventitious buds elongation for Pinus bangeana by adding GA3 in adventitious growing medium only. The rooting rates of Pinus ponderosa shoots increased from 2% to 15% by combing NAA and GA3 in medium.(7) The higher sucrose concentration could be an important factor for adventitious buds inducement and rooting of Pinus bangeana under the proper medium and hormone combination.(8) Activated carbon isn't good for adventitious buds inducement and differentiation, the adventitious buds growing can be promoted by adding 0.1% activated carbon in the medium, and the proper concentration of activated carbon can promote root growing.(9) The roots can be induced from Pinus ponderosa shoots at 1/2GD and 1/2SH medium. The roots inductive rate of Pinus ponderosa reaches 16.7% at 1/2GD medium with NAA 1.0mg·L-1 and GA3 0.5mg·L-1. The roots inductive rate reaches 8.3% at 1/2SH medium with NAA 0.5mg·L-1 and GA3 0.5mg·L-1, or with NAA 1.0mg·L-1 without GA3.