Study On Physiological And Biochemical Characteristics Of Organogenesis In Vitro Culture Of Ginger (Zingiber Officinale Rosc.)

Ginger(Zingiber officinale Rosc.)in this experiment were researched to establish,improve and perfect in vitro culture system as the root tip,stem apex and young leaf explants.In the process of the organogenesis,this work mainly focused on the research of biochemical indicators,soluble proteins and peroxidase isoenzyme,and further research on the morphogenetic callus has been done at the cytohistology level.The main results are as follows:1.Establishment of in vitro culture system in gingerThe result showed that different hormone played important roles in ginger organogenesis.The optimal method of explants sterilization was the 70%(V/V) alcohol to deal with 20 s,then the 5% NaClO to deal with 15 min.The optimal explant should be selected stem apex.The optimum ginger callus induction medium was MS + 2,4-D _{1.20 mg/ L} +6- BA _{3.0 mg/ L}.The optimum medium for the differentiation of budding ginger was MS +2,4-D_{1.20 mg/ L} +6- BA_{2.0mg / L} + NAA_{0.30mg / L}.The optimal seedling root tissue culture medium was MS + sucrose_{30g/ L}+ NAA_{0.30 mg/L} +6- BA_{3.0mg/ L} +0.03%activated carbon.The explants cultured in vitro selection methods should be solid - liquidsolid three alternate use of the medium.2.Researchs on physiological and biochemical indexesThe physiological and biochemical indexes in two kinds of callus are obviously different during the culture process.The physiological and biochemical indexes of the morphogenetic callus had wider change range than those of the non-morphogenetic callus.When the callus had been cultivated for eight days,the contents of nucleic acid(DNA,RNA),the antioxides enzyme(POD,SOD,CAT) activities of morphogenetic callus were higher than that of non-morphogenetic callus,and trends of those indexes were on the rise.At the same time,the contents of soluble sugars and starch were declined.While the changes of physiological and biochemical indexes of non-morphogenetic callus changed mildly.3.Researchs on soluble protein content and compositionThe average content of soluble proteins in morphogenetic callus was higher than the content in non-morphogenetic callus.Therefore,the lower-level of the proteins expression might be one of the important reason that let morphogenetic callus lost the capacity of redifferentiation and turned into non-morphogenetic callus.The results of SDS-PAGE electrophoresis results showed that morphogenesis of callus induction,proliferation and differentiation process of 10KDa,36KDa, 45KDa,55KDa,75KDa,85KDa,120KDa,180KDa,190KDa expression of the protein components.In the form of non-morphogenesis of the callus induction,proliferation and differentiation process 2KDa,10KDa,25KDa,36KDa and 70KDa expression of the protein components.â‘ only 10KDa protein molecular weight is shared between the two,shows that both callus induction in the early genetic material with the same expression.â‘¡36KDa protein in a callus morphogenesis patterns seen in the early stage of callus induction,and finally in the demise of differentiation,and in non-morphogenesis callus induction seen in the latter,there is not the demise of the last,that may be 36KDa protein so that the loss of differentiation ability of callus.â‘¢with callus morphogenesis map of 45KDa,55KDa,75KDa,85KDa,120KDa,180KDa,190KDa protein in the differentiation of adventitious buds no longer the expression of seven kinds of speculation is that this specific protein may promote the morph genesis of callus.4.Peroxidase isozymes ResearchPeroxidase isozyme analyse was as follows:the thickness of isozyme bands of the morphogenetic callus was little stronger than those of the non-morphogenetic callus.The morphogenetic callus had C and G bands that the non-morphogenetic callus didn't have.C band appeared after callus differentiation,so it could be used as a biochemical marker for callus differentiation.D band appeared during callus induction,this might be a sign for the begining of explants dedifferentiation.