The Establishment of Direct Regeneration System and Its Influencing Factors Using Petiole of Different Genotypes of Sugar Beet
|School||Chinese Academy of Agricultural Sciences|
|Course||Biochemistry and Molecular Biology|
|Keywords||Sugar beet Petiole Genotypic difference Regeneration system Browning Vetrification|
Sugar beet is an important economic crop. There has been much research on its tissue culture. A lot is about callus induction and bud differentiation, but relatively a little about direct regeneration of adventitious buds on explants. Acquirement of asterile sugar beet seedlings is first studied in the experiment. Then four genotypes, whose seedlings grow well, are preliminary selected from ten ones. By orthogonal design the best concentrations and combinations of plant growth regulator for adventitious bud induction and adventitious root induction of the four gentypes are selected, and the best prevention measures for browning and vetrification. At last acclimation time which influences survival rate is also studied. Direct regeneration system of different genotypes is preliminarily established, and browning and vetrification are prevented effectively. It prepares for genetic engineering breeding of sugar beet. The test results are as follows:1. Aseptic seedlings are acquired easierly from sterilizing sugar beet seeddirectly. In the course of peeling seed, scalpel cut is evidently better thancrandall knock in peeling effect.2. Screen out the best culture medium used for adventitious bud induction and induced rate:Culture medium: A、C:MS + 0.5mg/L 6-BA + 0.050 mg/L NAA B、D:MS + 0.5mg/L 6-BA + 0.025 mg/L NAA Induced rate: A:55.00%, B:42.50%, C:28.75%, D:37.50%3. Screen out the best culture medium used for adventitious root induction and induced rate:Culture medium: A、C:MS + 0.50mg/L NAA+ 0.50mg/L IBAB:MS + 0.25mg/L NAA+ 1.00mg/L IBA D:MS + 0.50mg/LNAA Induced rate: A:90.00%, B:87.50%, C:92.50%, D:100.00%4. The best prevention measure of genotype C browning: dark cultivation for 12 days, 3.0g/L Vc, 1.0g/L Ac. The rate of browning reduces to 8.33%.5. The best prevention measure of genotype D vetrification: 2000Lux light intensity, 8.0g/L agar, 40g/L sucrose. The rate of vetrification reduces to 8.75%.6. The best time for acclimation: 8 days. The survival rate is up to 96%.