Development Of The Device Of Mechanical Stress And Protoplast Isolation Of Callus In Echinacea Augustifolia

Plants play a significant role in ecosystem. Furthermore, it is an important reproducible source for human being. The ontogeny of every high plant as multicellular organism involves the following developmental processes: from a single cell to a mature individual, plant cells will undergo morphogenesis and differentiation. Nowadays, researchers pay more and more attention to these attractive phenomena. Plant biomechanics is an approach aiming at studying the interaction of the biophysics model and the environments, which are related to intro-cellular condition and extra-cellular environment. How to promote plant tissue growth is an attractive aspect in agriculture engineering and plant tissue engineering. The influence of mechanics factors on development and differentiation of single plant cell has been a potential method. In our research, three questions were proposed, how does plant tissue recognize environmental stress stimulate? Which effect do mechanics signals have on development and differentiation of plant cells? What signal transduction mechanism is? The correct model should be founded to predict the responses of pant cell signal transduction. Comprised the two key works, an effective experiment platform should be established to collect process data, and an efficient approach must be mastered to get enough plant single cells. To research the controlled biomechanics, the instrument of loading mechanical stress was developed. This sophisticated research instrument related to a device applying controllec mechanical forces to test specimens, particularly biological specimens, such as living tissue growing in cultures and single cell embeded in agarose. This study mainly intended to arrange for accurate manipulation of a specimen, and accurate measure of mechanical forces that it applied to, which proceeded in the light of basic theories, research means of biomechanics, mechanical design, circuitry, SCM, electromotor and plant physiology, etc. The instrument was a microprocessor-controlled mechanical loading system, specifically engineered for plant cell in sterile culture. The core of the control module of the instrument was SCM SPCE061A, and some chips in the circuit were used in it. The arrangement provided load between 50mN-300mN. A mass of plant single cells are needed in plant cell mechanics investigation. In this paper, Echinacea augustifolia from Canada was selected as material for protoplast isolation. The callus of Echinacea augustifolia were used for isolation and a orthogonal experiment was designed to research the factors influencing the process of protoplasts preparation, the result indicated that it was easy to isolate protoplast from buff-green Callus of Echinacea augustifolia which looked like granule with Symmetrical character. It was showed that the best enzyme solution concentration was composed of Cellulase 2.0% (w/v), Pectinase 1.0% (w/v), Hemicellulase 0.5% (w/v), 0.7mol·L-1 mannitol, and 50.0×104/gFW protoplasts were obtained after 8 hours later. Finally, we summarized the integrated approach including callus induction, callus culture, isolation, purification, identification and calculation of protoplasts.