Callus Induction And Plant Regeneration Of Eulaliopsis Binata Preliminary Study On Marking And Testing Of Cytoskeleton

The mature seeds of the apomictic plant, Eulaliopsis binata was used as the explant to study the callus induction, sub-culture, callus differentiation and the whole plant regeneration.The main results are as follows:1. As the same as other gramineous plants, the mature embryo of Eulaliopsis binata is sensitive to 2, 4-D but not the other plant hormones.2. The callus was induced successfully in a frequency of 97.27% in the MS medium plus with 2.5mg/L 2, 4-D, 8g/L agarose and 30g/L sucrose. Addition of proline and glutamine to the medium enhanced the calluses induction frequency to some extent, while ABA inhibited the callus formation. The sub-culture of callus also like this.3. The optimal medium for callus differentiation was MS+2.0mg/L 6-BA+0.1 mg/L NAA, the regeneration frequency reached 29.7%.4. We could not induce the callus from young leaf. The N6 and HB medium, which frequently used in gramineous plants, are not good for induction efficiency to Eulaliopsis binata.In addition to the maintenance cell shape, cytoskeleton also takes part in many important physiological activities. For the reason of plant cell inherent features, the labelling of plant cytoskeleton is still remaining some difficulties. In this research, we screen several ways of plant cytoskeleton labeling based on Loquat pollen. The results were showed:1. After fixation of the material, the non-cell bone proteins were taken away. Using Coomassie brilliant blue R₂₅₀ as dying regent, it could non-specifically bind with cell bone proteins. The distribution of cell bone proteins could be observed clearly in optical microscope. This result just similar with other researches.2. The phalloidin could specifically bind with microfilament. Using this feature, the fluorescein isothiocyanate-bind could be employed as marker. Using of fluorescent and confocol microscopes, we obtained the clear fluorescent image. The structure of microfilament was clear and brilliant.3. The microtube proteins were labeled with immuno-fluorescence technology. Combine by the fluorescence micro-technology, we could observe the microtube clearly.4. Comparing with these three methods, the results were almost the same. Coomassie brilliant blue dying was convenient, quick, easy-handle and had a low cost. The other two methods could detect the microtube and microfilament, but they were high cost and fussy procedures. For the specific binding, the results were more credibility. Furthermore, the clear image quality has direct meaning for some researches of physiology and biochemistry.