The Establishments And Optimizations Of In Vivo Cultutre And Regeneration System For Four Poplar Species

With the rapid growth of world’s population, increasing ecological environment destruction, coupled with the limitation of geographic and climate conditions, traditional breeding and breeding methods cannot satisfy the current situation, the resistance breeding of poplar also appears extremely urgent, and the basis and key point of resistance breeding is the establishment of efficient regeneration system of poplar. This study is devoted to use the Cicc Yang7、Populus simonii、Insect-resistant Yang2 and Populus bolleana to establish appropriate medium poplar leaf as explant regeneration system, to provide materials for study of poplar conversion system, laying a foundation for art poplar transgenic breeding.This study is to filter disinfection time screening of poplar leaves, petioles, and leaf segments of Cicc Yang7、Populus simonii、Insect-resistant Yang2 and Populus bolleanaYang,in order to select the most suitable explant and the best time to disinfect. We get the best differentiation medium formula through optimization of BA and NAA concentration ratio with sterile leaves as explant. We get the best rooting medium formula through selecting growth differentiation of buds and adjusting the concentration of IBA.Also, we get the best formula of the callus induction medium through adjusting of BA and NAA concentration ratio with sterile leaves as explant. The test results are as follows:(1) The most suitable material of Cicc Yang7 for tissue culture is petiole,with 75% alcohol solution 15s, 0.1% HgCl2 solution of 7min, pollution rate 3.3%, the rate of 96.7%. With Cicc 7 Yang aseptic leaves as explant, the best differentiation medium was MS+0.5 mg/LBA+0.5 mg/LNAA and MS+2.0 mg/LBA+ 0.5 mg/LNAA. The best rooting medium for 1/2 MS+0.3 mg/LIBA. The best medium for callus MS+0.5 mg/LBA+1.0 mg/LNAA and MS+0.5 mg/LBA+0.5 mg/LNAA.(2) The most suitable material of Populus simonii for tissue culture is petiole.with 75%alcohol solution 15s.0.1% HgCl2 solution of 8min. pollution rate 3.3%. the rate of 95.0%. With Populus simonii aseptic leaves as explant, the best differentiation medium was MS+2.0 mg/LBA+0.5 mg/LNAA. The best rooting medium for 1/2 MS+0.5 mg/LIBA. The best medium for callus MS+0.5 mg/LBA+1.0 mg/LNAA and MS+1.0 mg/LBA+1.0mg/LNAA.(3) The most suitable material of Insect-resistant Yang2 for tissue culture is stem and leave, with 75% alcohol solution 15s.0.1%HgCl2 solution of 7min, pollution rate 15%. the rate of 83.3%. With Insect-resistant Yang2 aseptic leaves as explant. the best differentiation medium was MS+0.5 mg/LBA +0.3 mg/LNAA and MS+2.0 mg/LBA+0.5 mg/LNAA. The best rooting medium tor 1/2 MS+0.5 mg/LIBA. The best medium for callus MS+1.0 mg/LBA+1.0 mg/LNAA.(4) The most suitable material of Populus bolleana for tissue culture is petiole,with 75%alcohol solution 15s,0.1% HgCl2 solution of 8min, pollution ratel0%, the rate of 90%. With Populus bolleana aseptic leaves as explant, the best differentiation medium was MS+0.5 mg/LBA+0.5mg/LNAA. The best rooting medium for 1/2 MS+ 0.3mg/LIBA. The best medium for callus MS+ 0.5mg/LBA+ 0.5 mg/LNAA and and MS+ 1.0 mg/LBA+ 0.5mg/LNAA.