Analysis On Green Leaf Mutant In Dioscorea Alata.L

Dioseorea alata L. belongs to Dioscoreaceae Dioscorea vine.It is high starch content, especially its rich anthocyanin loved by the people. Anthocyanins have anti-aging,anti-oxidation,anti-tumor,prevention and treatment of cardiovascular and cerebrovascular diseases, lowering blood sugar,anti-inflammatory,and promote visual and so on.now domestic and foreign pharmaceutical and food sector widespread attention.In this study, select the Dioscorea alata L.from the Dioscorea resource garden of college of Agriculture Hainan University as the material. Study on effect of Ca2+on callus induction of Dioseorea alata L. and identify the type of callus organization by paraffin section.Initially establish a chemical mutagen EMS (Ethylmethylsulfone), NaN3(Sodium azide),DES(Ethyl sulfate), and mutagenic repair inhibitor benzamide treatment embryogenic callus of methods and techniques. Study on gene cloning and analysis of by reference to the Dioseorea alata L.anthocyanin synthetic of critical sites of explain the the authenticity of the the DNA variation.Effect of anthocyanin synthesic about physiological parameters was measured.Analysis of caused by natural plant causes color change under culture conditions. The main conclusions obtained are as follows:1. Study on effect of Ca2+on callus induction of Dioseorea alata L. Results showed that:with elevated concentration of Ca2+explants out more taken the lead in reducing elevated,3×Ca2+out the best callus induction rate of33%. The callus forms loose type into dense type.2. Study on establishment of embryogenic callus induction system:Stem were inoculated in the hormone-free MS medium, cultured for60days.About1cm of section stem inoculated into MS+0.2mg/LNAA+1.0mg/L6-BA+3×Ga2+medium, and cultured for40-50days;selected node at the granular tissue,divestiture the granular tissue with a sterile forceps and sterile blade,in this case, the callus is transferred to MS+0.2mg/L NAA+1.0mg/L6-BA medium resumed training about two weeks.3. Through that the tissuel paraffin section. Results there are obvious embryonic tissue cells, some organizations also form embryoid bodies, a part of the organization showed a cell divided into two developmental phenomena.the organization proved a single cell origin, can be initially identify by the organization to embryonic callus.4Establish a chemical mutagen EMS, NaN3, DES, and mutagenic repair inhibitor benzamide treatment embryogenic callus of methods and techniques. Optimal mutagenesis conditions:first choice3mM of sodium azides treated under pressure forlhours, and then choice5mM of sodium azides treated under pressure for4hours.The processing conditions of27℃,100r/min.5. cloning and anthocyanins variant analysis and studies of light intensity on anthocyanin synthesis of findings:anthocyanin gene variant partial fragment of occurred the individual nucleotide substitution; light intensity on the synthesis of anthocyanins have a significant impact, so as to analyze the occurred leaf variants variation may be due to changes in the base level, light intensity, but a supporting role...