| Black locust (Robinia pseudoacacia L.) is a multipurpose legume tree species. It has fine characteristics of resistance to drought, salinity, and is one of the Pioneer Tree Species for land reclamation and reforestation in China.In addition, it is also the well-known gardening trees and ornamental trees. Tetraploid black locust is a new varieties which is not only has the fine characteristics of the black locust, also has the fine features of large leaves, high protein contents and so on.In this test, the effect of different plant growth regulators and the number of subculture on callus induction and cell suspension culture and plant regeneration were studied by using cotyledon explants which had grown7days from black locust seeds. The results showed that different types and concentration of the plant growth regulators had influenced on callus induction and adventitious bud regeneration from callus of black locust cotyledon, the best culture rate of plant growth regulators of inducing callus from cotyledon and the best culture medium of suspension cells both are MS+2,4-D1.0mg/L+6-BA0.2mg/L, callus induction rate is98.9%;the suspension cell liquid was yellow, and small particulate were got. The medium of adventitious bud induction from callus is MS+6-BA1.0mg/L+NAA0.05mg/L, adventitious bud induction rate is74.4%. Adventitious bud induction frequency of callus declined as the subculture number was increased. The highest adventitious bud induction rate that was induced directly from the black locust cotyledon was87.8%, and its medium was MS+6-BA5.0mg/L+2,4-D0.1mg/L.Orthogonal design was used that three growing stages of diploid locust seeds was soaking in colchicine. The colchicine concentration was0.1%,0.3%,0.5%, and the processing time is for1,2,3d respectively. The results showed that using the concentration of0.3%colchicine to soak the black locust seeds which was burst for three days. Both higher survival rate and polyploid induction rate of black locust seedlings were obtained. |
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