Studies On The Germplasm Rejuvenation Of Dendranthema Morifolium (Ramat.)Tzvel.cv.Gongju

Dendranthema morifolium (Ramat.) Tzvel. cv. Gongju belongs to the perennial herbaceous of Compositae. The roots, stems, leaves and flowers all can be used as medicine, It can cure the liver eyesight, detoxification swelling and other effects. It is one of the authentic ingredients of Anhui Province, and can be used to cure head wind, dizziness, Longbitong, sore throat, carbuncles boils drugs, and other symptoms in clinical practice. It has been a long time since Dendranthema morifolium (Ramat.) Tzvel. was used as the raw material medicine in the proprietary system, and the effect is significant, the demand of market for it is increasing number, such as Qijudihuang pills, tablets Sambo, heat eyesight tea, etc.In the large-scale production, it mainly uses asexual reproduction such as cuttings and ramet. But in that way, Dendranthema morifolium (Ramat.) Tzvel. is vulnerable to a variety of viruses and other microbial attack; which being infringed infected for life, over the cultivation time, the disease would worsen, that will lead to production cuts, germplasm degraded,the more serious consequences is the plant died; The physical, chemical and other control measures effect for the prevention and treatment of viral diseases is not significant.Objective:The purpose of the test is to establish an in vitro rapid propagation system, and rejuvenation the germplasm of Dendranthema morifolium (Ramat.) Tzvel. cv. Gongju, in order to ensure the genetic stability of plants; and rejuvenation the germplasm of Dendranthema morifolium (Ramat.) Tzvel. cv. Gongju, in order to ensure the genetic stability of plants; lay the foundation for the standardization of high quality seeds and seedlings (GAP) industrial production.Methods:This test is mainly through the in vitro shoot tip culture of Dendranthema morifolium (Ramat.) Tzvel. cv. Gongju which is improved varieties introduced to eastern Guangdong Province. Plantlets obtained using heat treatment combined tip culture method to remove the virus in order to rejuvenation of its germplasm, and use the way of axillary bud breeding to build the vitro propagation system of Dendranthema morifolium (Ramat.) Tzvel. cv. Gongju to ensure the genetic stability. The tests conducted mainly through literature research, field surveys, single factor experiment and orthogonal design method.Results:This study summarizes the research status of Dendranthema morifolium (Ramat.) Tzvel. germplasm, the content covered the research progress of herbal research, research pharmacognosy, chemical, pharmacological, clinical application, biology, detoxification methods and identification, tissue culture and other aspects of Dendranthema morifolium (Ramat.) Tzvel.. Through the experimental study, we screened out the best medium and cultivation techniques of primary culture, subculture, root seedlings, domestication and transplanting etc. phases, optimize and improve the tissue culture system of Dendranthema morifolium (Ramat.) Tzvel. cv. Gongju:This test established detoxification techniques and identification method which is suitable for industrial production,which is strong feasibility, low-cost and the effect of virus removal is remarkable.Conclusion:1.The establishment of tissue culture system about Dendranthema morifolium (Ramat.) Tzvel. cv. Gongju.1)The acquisition of sterile tissue culture seedlings:the more appropriate disinfection solvent is 70% ethanol and 0.1% mercuric chloride, for the shoot tip explants, the desired effect is 70% alcohol sterilization 45 s then sterilized with 0.1% mercuric chloride about 3 min; for stem segments, the desired effect is 70% alcohol sterilization for 30 s and then sterilized with 0.1% mercuric chloride for 11 min.2)Axillary bud induction:In this experiment, MS as the basic medium, the plant growth substances which have the better inducing effect are cytokinin 6-BA and growth hormone NAA, The culture medium MS+6-BA1.5-2.0 mg/L+NAA0.1 mg/L is conducive to the formation of axillary buds, after 45 days culture, the proliferation multiple is 5.3 to 5.8, and the axillary buds growing well.3)Various effect of additives on the growth of tissue culture seedlings:it can significantly promote the growth of tissue culture seedlings when the amount of macronutrients was 150 mL/L, the amount of trace elements is 1 mL/L and the vitamin dosage is 1.5 mL/L; More appropriate agar concentration was 4 g/L, coconut milk is the preferred natural additives, and in the concentration range of 50-200 mL/L, the tissue culture seedlings growth accelerated as the concentration of growth, expressed as height increased gradually, stems relatively stout, leaves dark green and leaf increases; According to the results of the orthogonal experiment, for tissue culture seedlings, the most beneficial combination of medium for seedlings is 1/2 MS+sucrose 30 g/L+AC 1 g/L+NAA0.1 mg/L+IBA0.1 mg/L, in which, sucrose on tissue culture seedlings grown the greatest impact.4)Rooting induction culture:the medium of 1/2MS+IBA 0.05 mg/L+NAA0.05 mg/L is suitable for rooting; Adding activated carbon is conducive to the lateral roots growth of tissue culture.5)The program of acclimatization:the plantlets cultured about 30 days in rooting induction medium until tissue culture seedlings grown to more than five effective leaves, then opening the lid about five days and then hydroponics three days to ensure tissue culture seedlings to adapt to the environment, finally, planted in vermiculite:sand (1:1)2. Virus removal and identification of tissue culture seedlings.This study using a combination of heat treatment and secondary tip culture in detoxification methods and trait identification combine indicating plant identification in the identification of virus-free, In the long-term subculture, tissue culture seedlings which has not been detoxification treatment exhibited leaf yellowing, black spot, leaf, shrink the top and other symptoms, however, virus-free seedlings did not show the corresponding symptoms, and all are growing well, the tissue culture seedlings without detoxification treatment make cowpea showed chlorotic spots, then death after the virus spread to the entire plant, however, the cowpea plants growth healthy in the virus-free seedlings Group.