Tissue Culture, Photosynthesis And MiRNA Characterization Of Typhonium Flagelliforme (Lodd.) Blume.

Typhonium flagelliforme (Lodd.) Blume., a member of the family Araceae, which has been used as a traditional Chinese herb over thousands of years. The objective of this thesis was related to analysis of tissue cluture, photosynthesis characteristic and miRNA deteting.Furthermore, the first part of this thesis was to build up a rapid propagation method of T. flagelliforme for the purpose of large-scale breeding and high quality of seedlings. Finally, a standard method of tissue culture was set up. Asexual propagation system of the tuber tissue was established using mediums with different kinds of hormones. Firstly, the constituent of callus culture medium was prepared as following: MS + NAA 0.2 mg/L + KT 1.0 mg/L + sucrose 3 % + agar 6 g/L, and callus of T. flagelliforme can be generated successfully. Secondly, Medium was optimized to generate adventitious buds efficiently, especially for higher proliferated multiples, which consists of MS + NAA 0.2 mg/L + 6-BA 2.0 mg/L + sucrose 3 % + agar 6 g/L. Eventually, medium consisted of 1/2 MS + NAA 0.2 mg/L + IBA 0.4 mg/L + KT 0.2 mg/L + sucrose 3 % + agar 6 g/L + AC 0.3 g/L was beneficial to roots generating, and regenerative plants can be obtained 12-14 weeks later. After experiment of one-step-seedling formation was carried out, it was indicated that medium containing MS + NAA 0.2 mg/L + 6-BA 1.5 mg/L + sucrose 3 % + agar 6 g/L was the best of all, and new plantlets could be developed 5-6 weeks later, seedlings adaptation can be carried on 10 weeks later.The growth, photosynthetic gas exchange and chlorophyll ?uorescence were studied in T. flagelliforme infected with cucumber mosaic virus by using photosynthesis analyzer, providing evidence for control of virus disease. The infected plants showed decreased plant weight, leaf area, plant height although they had similar leaf number. Decreased net photosynthetic rate (Pn), transpiration (E) and stomatal conductance (Gs), and a slightly higher intercellular CO2 concentration (Ci) were observed in infected plants. Chlorophyll content was significantly reduced infected plants as compared to mock, which suppressed the plant growing directly, and caused the negative influence. In conclusion, T. flagelliforme after CMV infection showed a general response as reported in other plants infected by virus.Finally, miRNA types and number of T. flagelliforme were studied through real-time PCR and micro fluid chip (microarray) technology, and the gap of miRNA of T. flagelliforme can be filled forever in Sanger miRbase. On one hand, existences of 10 miRNAs were verified through realtime-PCR, but the result showed that genes accumulation was different. On the other hand, 734 miRNAs were detected out of the original designed 1450 probes of mature miRNA in 34 kinds of plants, which account for 50.62 %, distributed in 317 miRNAs family. It is demonstrated that the miRNA distributed generally and the conservatively. Original intention of this experiment was to confirm the result between real-time PCR and chip, however, it was contradicted. Further analysis of the reliability and accuracy of two methods above needs more experiment in the next research.