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The Construction of Suspension Culture and Primary Study on the Mechanism of Stress Resistance of Capparis Spinosa L.

Author: LiuZuo
Tutor: SuMaoTeng
School: Huazhong University of Science and Technology
Course: Biochemistry and Molecular Biology
Keywords: Capparis spinosa L. Cell suspension culture Physiology and biochemistry Environment stress Volatile oil
CLC: S567.19
Type: Master's thesis
Year: 2009
Downloads: 54
Quote: 0
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Abstract


Capparis spinosa L. is a very important plant in ecological and economical effects. However, the narrow distribution and the gradually decrease of resources restricted its’application. Plant cell culture is regarded as a promising way to provide an attractive alternative source for limited natural resources.This paper mainly focused on physiological and biochemical characters of different sizes of C. spinosa cell aggregation, metabolic process of nutrition compounds in cell suspension culture, physiological response of C. spinosa to water stress and low temperature, and the content of volatile oil in fruit and seed of C. spinosa. The main results were shown as follows:(1) According to the diameters of the cell, we selected three different sizes of C. spinosa cell aggregation and determined flavone content, correlated enzyme activities and recultivate curve. The results indicated that the flavone content of the smallest cell aggregation was approximately twice compared to the larger ones, and its malonaldehyde content (MDA), the phenylalanine ammonialyase (PAL) and peroxide enzyme (POD) activities and recultivate curve presented the same tendency. The small cell aggregation contacts fully with the oxygen and the nutrient of culture medium, which is beneficial to cell growth and the secondary metabolite accumulation.(2) The cell growth in suspension culture and metabolic process of nutrition compounds in medium were investigated. It was showed that the pH value of suspension cultures fluctuated between 4.2 and 5.8. It was showed that the pH value declined in the beginning and the rise. Phosphate and nitrate met the needs basically. Phosphate in medium was ingested quickly, followed by NH4+ and NO3-. The content of soluble cellular protein rised at first then dropped, and it had a linearly correlation with the content of extracellular protein.(3) The effect of water deficit on physiological parameters was studied in celll suspension culture of C. spinosa to investigate its drought tolerance. The results indicated that with the increased PEG-8000 concentration (4%-10%) and prolongable time, the content of MDA rose, and the content of Pro increased first and then decreased. As for the content of Pr and the activity of POD, they experienced a increased process, and total unsaturated fatty acids went up. This demonstrated that the membrane deroxidization of C. spinosa was stronger, and the system of cell defense enzyme took action again the damage caused by water stress.As the PEG concentration increased (above 10%), there was a slight decrease in IUFA which indicated the excessus damage towards cell membrance.(4) The physilological-biochemical effects of temperature stress on C. spinosa was studied, and we conducted that damage of cell membrance occurred, inducing the content of MDA and the activity of POD increased. Besides that, the content of soluble sugar, protein and free proline had distinct increased, and total unsaturated fatty acids went up, which were instrinsic factors to improve repelling temperature stress.(5) GC-MS method was established to measure the content of volatile oil in fruit and seed of C. spinosa. The results suggested that the main elements included undersaturation acids, alkane, Phenols and esters, with undersaturation acids taking the largest percentage.

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