| Photoautotrophic micropropagation is new technology of plant tissue culture created by Japanese Professor T.Kozai from Chiba University in Japan, a solution to the defects found in the traditional techonlogy of tissue culture, i.e. vulnerable contamination, variation, poor growth, poor rhizogenesis and a higher mortality during periods of acclimation and transplantation. The technology is typically applied to seedling production, which is featured by controlled conditions, CO2 as a substitute for sugar as a carbon source, by means of providing optimum light, temperature, water, air and nutrition for the plantlets to boost their asphotosynthesis so as to promote their growth.The research on forest-tree photoautotrophic micropropagation was carried out under the instruction of Professor T.Kozai, and based on the equipments for photoautotrophic micropropagation developed independently previously (two related patents are being applied, whose numbers are 20071004506.X and 200810071155.9 ). Aimed at the long cycle in forest-tree seedling breeding and optimization ineffeciency in environmental parameters, the research was to explore a new method of parameter optimization with eucalypts as test materials by applying the computable three stages design widely used in industry to photoautotrophic micropropagation. At the stage of design, a simplex method was used to optimalize the parameter combination of physical environmental factors. In addition, using chlorophyll fluorescence imaging and root scan systems, the effectiveness of parameter optimization was also studied and the effectiveness of the three stage design applied in the research was tested.1. Aimed at the features of forest-tree seedling breeding, a set of photoautotrophic micropropagation system was developed in the research, including system of both hardware and software, where both liquid-phase culture and solid-phase culture are available. There into liquid-phase culture by means of atomization greatly boosted the dissolved oxygen in culture solution. The software system with remote supervision function includes the client and the sever.2. On the basis of Window XP, C++ builder 6.0 and SQL Server 2000 and in combination with software components of TIMER and SERIALPORT, the client was a set of client management system for forest-tree photoautotrophic micropropagation, which is characterized by more security, stable performance, good portability and great convenience to fully meet the control requirement.3. Based on Window XP, C++ builder 6.0 and SQL Server 2000 and combined with software components of TCLIENTSOCKET and TSEVERSOCKET, the sever was a set of sever management system with emote supervision function for forest-tree photoautotrophic micropropagation. Many advanced features plus the design philosophy for the graphical user interface of Microsoft Windows were applied to the system, that is, the system made it possible for several clients to retrieve, update or delete data from the same database table while keeping the consistency and integrality of the data with clients unnecessary to wait. Furthermore, the system also has such advantages as more security, stable performance, fast speed, friendly interface and good portability.4. With eucalypts as test materials and by applying the computable three stage design(system design, parameter design, and tolerance design), the research was to explore optimization of four major physical environmental factors, humidity, temperature, light intensity and density of CO2, which affected forest-tree photoautotrophic micropropagation.1) At the system design stage, by applying quadratic regressive orthogonal design to the research, established was the control model of major physical environmental factors (humidity, temperature, light intensity and density of CO2), measured by rooting rate.2) At the parameter design stage,parameters were designed by using a revised simplex method through 12 trials altogether, which resulted in the optimal response of rooting rate, about 93.96%, the condition where the humidity, temperature, light intensity and density of CO2 was 125.00μmolm-2s-1, 1500.00ppm, 24.00℃and 75% respectively.5. Parameter optimization had a positive effect on chlorophyll fluorescence parameters, however the effects on Rfd,Fm', and Qy_max were only significant, while for other chlorophyll fluorescence parameters, not significant. Therefore, to test the effectiveness of the optimization, chlorophyll fluorescence parameters are supposed to be measured by Rfd,Fm', and Qy_max.6. Parameter optimization had different effects on different root morphological indexes. Different physical environmental parameter combinations had significant effects on the overall length, tip numbers, and crossover-point numbers of eucalypt seedlings'roots through photoautotrophic micropropagation, while the effects on their projected area, surface area, mean diameters and overall volume were not significant. Therefore, to test the effectiveness of the optimization, root morphological indexes are supposed to be measured by the overall length, tip numbers, and crossover-point numbers.The dissertation was to explore new theories and technology of forest-tree seedling breeding and was also an applied research on new methods of photoautotrophic micropropagation. Not only did it lay the foundation for forest-tree photoautotrophic micropropagation, but also enriched and developed the theories of forest-tree seedling breeding. It can reduce the production cost of tissue culture seedlings, shorten the cycle of improved variety promotion, improve the quality of seedlings and greatly improve the industrialized seedling production. Meanwhile, it can also solve the contradiction between supply and demand of superior seedlings and meet the market demand; therefore it is of vast potential in the future market.
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