| Energy crisis is a enormous challenge that the human will have to face, bio-energy development and utilization has become a hot topic. Energy plants can be directly or indirectly as fuel instead of fossil fuels, so in order to improve Datura stramonium seed oil content, This article which as its hypocotyl explants, initially established Datura stramonium tissue culture system, providing the basis for transgenic of training for high oil content; In addition, We also determine 4 species of seed oil content using two different methods, and compared the two methods, in order to find potential energy plants to provide the scientific method.The main conclusions of this paper as follow:1. Soaked with 70% alcohol sterilization for 30s, then soaked for 20min in 30% sodium hypochlorite solution, Rinsed with sterile water and inoculated into MS medium without hormone; take sterile seedlings which of seedling age is 5-7 days, cut its hypocotyl to 0.5-1.0cm in subparagraph, To callus induction medium inoculated, dark culture for 48 hours then transfer into incubator; when callus diameter growth to be about 1.0-1.5cm, removed and transferred to shoot induction medium for induction; when the variable shoot grow to about 3-4cm, Go to rooting medium for Culture(Culture conditions are all temperature of 26±1℃, humidity 70% -80%, light 14h·d-1, light intensity 2500lux). Callus induction medium of hypocotyl is 6-BA 1.0 mg·L-1 + 2,4-D 0.3 mg·L-1, induction rate is 96.67%; Shoot induction medium is the 6-BA 2.5 mg·L-1 + NAA 0.1 mg·L-1, the adventitious buds is 57.78%;, the best rooting medium is 1/2MS + IBA 0.2 mg·L -1 + NAA 0.05 mg·L-1, the rooting rate is 60.00%; hardened after transplanting, the survival rate of tissue culture is 90%.2. After different treatment of Datura stramonium seeds, the germination rate and germination potential have occurred major changes. Soaking seeds in 55℃and 80℃hot water achieves good results, with the germination rates are 40% and 47.5%, germination potential are both 35% ; However affter soaking seed with concentrated sulfuric acid, the germination rate and germination potentia reached the highest , which is 67.5% and 42.5%,respectively. The resulting indicate that the best treatment to raise Datura stramonium seed germination rate and germination potential is soaking in concentrated sulfuric acid for 20min and in warm water of 45℃for 24h.3. we measured the seed oil contents of four potential energy plants using the methods of Nuclear Magnetic Resonance (NMR) and Soxhlet Extraction, and compared the results between them. Results showed that the seed oil contents tested by NMR are significantly higher than that of Soxhlet Extraction. Our data indicate that NMR method has advantages of simple and no damage, which avoids the disadvantages of Soxhlet Extraction such as time-consuming and difficult extract fatty acids with combination state (e.g. Lecithin, Glucolipid, Lipoprotein, etc.). Thus, NMR can be used for rapid testing of seed oil content, which provides a reliable method for screening potential energy plants. |
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