Study On Organogenesis Regeneration And Genetic Transformation Of Sugarbeet In Vitro

Sugarbeet is an important sugar and forage crop, all over the world, sugarbeet as sugar provides about 30 percent sugar for people. Sugarbeet as forage is best succulent feed for domestic animals. Its earthnut is weighty, and its nutrition is abundant. Because of cultivated and domesticative time for sugarbeet is short and it is biennial plants and xenial crop, genetic background is straitness and resource is scarce. Especially, what self-crossing is highly no appetency makes easy lose of good single genotype, which propagatation and conservation of good varieties breeding brings great difficulty. The developmnet of biotechnology bring a new path for sugarbeet breeding. But because repetition of sugarbeet tissue culture is low, callus induction is difficulty, regeneration in vitro is badly genetype-dependent, aggravates localization of straitness of variational begats and genetic basis of breeding and planting varieties, which lead to sugarbeet industry in our country to be faced with baptism and planting areas largely decreases in competition with sugarcane sugar.As the problems narrated above, the study is on the transformation and the test of anti-virus genes transplanted to six different cultivars, and on the analysis of isoenzymes level in different developmental stages.1 Shoot induction of sugarbeet cotyledon nodes is genetype-independent.With these six cultivars, when cotyledon nodes were cultured on multiple shoots induction medium, the results were not appeared much difference between these cultivars. It is showed that multiple shoots induction was not under the control of different genotype.2 While 100mg/L Kan and 500mg/L Cb were added into the selective medium, high transformation frequency can be obtained. When infection,washing with steriled water adding 800mg/LCb. In this research, 100mg/L AS was used to activate gene and enhance transformation rate.3 Shoot induction from cotyledon nodes was under different combination of BA and NAA. High concentration of BA could get more shoots inducted from cotyledon nodes ,but some vitrification included appeared at the same time. When cut off the leaves, we got the healthy shoots from the stem nodes. Among these six cultivars, the vitrification of 51130 and 51132 were more serious than other three.4 Multiple shoots induction from leaf and petiole was restricted by different genotypes, different cultivars get distinct multiple shoots induction rate. However, it made much difference when use smaller shoots for shoots proliferation, it didn't effected by genotypes.5 Root development initiated in about 10days and got maturation after 20days. Root system is thick and vigorous.Single shoot for rooting with MS dedium adding 3~5mg/l IBA and 300mg/l Cb.6 PCR test of transformed plantlet is positive. The band of RF and PBVP6 genes is clear-cut on PAGE lane. It is showed that RF and PBVP6 were sucessfully transplanted to sugarbeet.7 The paper also dealt with the study on the peroxidase and esterase isozymes as well as change of soluble protein content in the process of direct shoots organogenesis and genetic transformation of sugerbeet in vitro by polyarclamide gel electrophoresis (PAGE). The results showed that peroxidase (POD), isoesterase (EST) isoenzyme and soluble protein content appeared the changes of enzyme zymogram number and colour shade by adding benzylaminopurine to the medium, which related with increase of metabolizing activity. And after genetic transformation, bands of the peroxidase and esterase isozymes and soluble protein came forth obviously colour darken and number increase, which do further research to identify the resons that resistance to stress was enhanced or natural transgene.