Study On The Plant Tissue Culture And Plant Cell Fusion Of Mozzie Buster And Ocimum Basilicum L.

Mozzie buster is cell fusion product of Africa's Geranium and China'sCymbopogon citrates. It is a kind of aromatic perennial herb plant. Citrosa can bedistributed by its leaves to have a good insect repellent effect. Besides, it also have theeffect of refreshing, antibacterial, antidepressant, easing the pressure and is praised as"the god grass". Because Mozzie buster is somatic cell hybrid products, seed sterility,and the conventional method of cutting the rate of reproduction is low, therefore vitrorapid propagation technology is the practical technological means. Ocimum basilicumis not only raw materials of perfume, but also has fly-repelling effectiveness. Thedevelopment of biotechnology has made it possible to breed a new variety of Mozziebuster which can repel both mosquito and fly. In this study, the whole tissuecultivating system of Mozzie buster and Ocimum basilicum was established, and thecondition of protoplasts fusion was primarily analyzed. The results of our experimentwere shown as follows:1. MS was used as the basic cultivating medium. Based on taking the orthogonaldesign experiments and by using the technique of plant tissue culture in this paper, theexperiment-system of tissue culture and rapid propagation of Mozzie buster andOcimum basilicum have been successfully established. The optimal cultivatingconditions of Mozzie buster wsa: the most suitable medium for the initiation ofadventitious buds is MS+6-BA0.2mg·L^-1+NAA0.3mg·L^-1; The best mediurn for therooting of shoots is 1/2MS+NAA0.4mg·L^-1 and the optimal medium for theinducement of callus tissues is MS+2,4-D0.5mg·L^-1+6-BA 2.0 mg·L^-1+NAA0.3mg·L^-1. The optimal cultivating conditions of Ocimum basilicum was: themost suitable medium for strong seedling is MS+6-BA0.2 mg·L^-1 +NAA0.2mg·L^-1+KT0.3mg·L^-1; The best medium for the rooting of shoots is 1/2MS and theoptimal medium for the inducement of callus tissues is MS+ 2,4-D 0.5mg·L^-1 +6-BA3.0mg·L^-1+ NAA 0.5mg·L^-1.2. The best combination of isolating protoplast of Mozzie buster was the enzyme mixture containing 2.0% Cellulase R-10, 0.4% Pectinase, 0.5M mannitol as osmoticpressure, 27℃under dark 80r/m rotating enzymolysis 8h, and the highest yieldachieved to 1.67×10⁶per gram, the protoplast vitality was 87.4%; The bestcombination of isolating protoplast of Ocimum basilicum was the enzyme mixturecontaining 1.6% Cellulase R-10, 0.4.% Pectinase, 0.5M mannitol as osmotic pressure,27℃under dark 80r/m rotating enzymolysis 8h, and the highest yield achieved to1.67×10⁶ per gram, the protoplast vitality was 87.4%.3. By studying the method of PEG induced protoplast fusion research, the bestconditions for integration of Mozzie buster and Ocimum basilicum was30%PEG(6000) with 0.3 mg·L^-1 Ca²⁺ and pH 10.5, and the protoplast fusionpercentage was 11.2%.