Studies On Plant Open Tissue Culture And New Propagation Technique

The experiment was done in Shandong Agriculture University, AriCompany of Jinan and Jinqiu seed Company of Shandong from 2003 to 2005.As for the questions such as complicated process, high contanmination rate,high cost and low living rate, based on the plantiotics, a new tissue culture systemwas set up , steps of which were simplified radically and the cost was reduced greatly.Based on the new system, the system of propagation technique was optimizedand adjusted, and became more effective and practical. The main results are asfollowing:1. Research of the plantiotics and experiments on effect1.1 Using the theory of traditional Chinese medicine, an active substancenamed as plantiotics, was abstracted from several medicinal herbs. Plantioticscould kill all kinds of bacteria and fungi broad-spectrumly.1.2 The experiment on fungoids falling naturally and artificial showed thatplantiotics could kill the spallogenic fungoids effectively in 0.3% and 1.9% forfungoids masses, above 10% can hurt the plants.2. Built of open tissue culture techniqueBased on the plantiotics, all steps of tissue culture were simplified radically anda new open tissue culture mode was set up successfully :2.1 Choice of the culture container: without autoclaving, choose the one-offplastic cups sealing with PE preservative film.2.2 Preparation for culture medium: when the agar is boiling, add sugar, all kinds ofnutritive elements, hormone and plantiotics instead of autoclaving, then adjust PH to5.6-5.8. Distribute the medium to plastic cups and seal them with PE preservative filminside an hour.2.3 Sterilization of the inoculating instruments: dip them in 20% plantiotics for an hour,the instruments being used should be dipped in 20% plantiotics during the whole periodof inoculating.2.4 Inoculating and culturing: the inoculating room should be sterilizated with 75%alhocol and ultraviolet lamp. Uncover the dime of preservative film, insert the plantmaterials in medium with inoculating instruments, then cover the cups quickly. Put themin the culturing room or greenhouse keeping light 1500-4000Lux and the temperature25℃. During culturing, keep the entironment clean and pick up the contaminations intime.3. Effect of the open tissue culture Aiming at the different period of open tissue culture, we had done a lot of contrastexperiments and conformed the best concentration of plantiotics.3.1 Aiming at the soaking concentration, soaking time and culturing concentration ofplantiotics, we had done lots of experiments on establishment of varieties of explants. Theresults showed that the success rate of explants could increase to above 75% from 30-40%traditionally, some could arrive 95%. Activated carbon could absorb the plantiotics, madeit losing the ability of killing bacterias and fungies. In order to prevent browning, we hadto use PVP or vitamin C instead of activated carbon.3.2 In differentiation period, the best concentration of plantiotics in medium was 0.5-0.7%.The differentiation rate was normal and the plants grew strongly. One reason was thatwithout autoclaving only a little of the nutritive elements and hormone was lost. Anotherwas that the light transmittance of plastic cups was better than glass container.3.3 In rooting period, the best concentration of plantiotics in medium was 0.7%. The rootsand plants grew strongerly than traditional method. One reason was that withoutautoclaving only a little of the nutritive elements and hormone was lost. Another was thatthe light transmittance of plastic cups was better than glass container.3.4 Because the light transmittance of plastic cups was better than glass container and theroots grew stronger, the adaptability to the entironment of greenhouse was better and theliving rate of transplanting was higher than traditional method.4. Improvement of the culturing method In greenhouse, without using lamps the temperature could be kept 25℃and the lightcould be kept 1500-4000 Lux by using sunshade. So the plants could grow better ingreenhouse. Culturing in greenhouse, a lot of electricity and manpower energy were savedand the work efficiency was improved greatly. As a result, the cost of tissue culture was...