Analysis Of Differentially Expressed Proteins Upon Sporangium Development From Dryopteris Fragrans (L.) Schott

Dryopteris fragrans(L.)Schott is Dryopteris Adans Dryopteridaceae,which is mainly distributed in China’s Heilongjiang Province,Wudalianchi area. Country folk prescription records of Dryopteris fragrans can treat various skin diseases,in recent years,more and more people aware of important medicinal value of Dryopteris fragrans.Fern sporangium is an important place to produce spores,plays an important role in sexual reproduction,but it is not clear for details on this process,proteomics of fern sporangium and spore forming process in the sporangium has not been reported.In this research,it was divided development of the Dryopteris fragrans sporangium into three periods,period A which was young spore leaf by circinate state to the full expansion;period B which was Dryopteris fragrans sporangium formation and from white to pale yellow;period C which was mature sporangium,become dark brown respectively.We determined part of the cell protective enzyme activity,chlorophyll fluorescence parameters and analysis differential protein expression and distribution characteristics of the three stages through the experiment.The mainly results included:(1) peroxidase (POD) activity,malondialdehyde (MDA) and proline (Pro) content showed the same variation at different developmental stages,and had the highest content or activity in growth period B,the lowest content or activity in development period A,and had significant difference in3different developmental stages;superoxide dismutase (SOD) was the lowest in the development period of A,with the developmental process,the activity of SOD increased gradually,and increased significantly to the maximum value during the period B,then decreased slightly,period A had significant difference with B and C,and difference between B,C time was not significant;catalase (CAT) activity was the lowest fertility in period A,the activity of CAT increased gradually with development process,and increased significantly to the highest value during the period C,the period A,B and C had significant differences,differences between period A and B was not significant.(2) Chlorophyll fluorescence parameters showed:the development period B of Dryopteris fragrans sporangium had higher conversion efficiency of light energy,the maximum fluorescence parameters were increased significantly to the period C,but the energy conversion rate was decreased notably,in addition,the initial fluorescence was more stable during the development period C. The system of two-dimensional electrophoresis was improved,phenol-method was more suitable to extract the fern protein samples,two-dimensional electrophoresis of protein sample quantity of protein containing1500μg protein sample liquid effect was better.(3) The experiment of the sample was used the improved two dimensional electrophoresis system,and the electrophoresis pattern was obtained by analysis of protein spots,which showed Dryopteris fragrans sporangia development related proteins differentially expressed spots were41,it carried on the calibration and classified analysis isoelectric point and molecular weight showed that along with the development process always up-regulated protein were23,down regulated the expression of protein were5,down regulated after up-regulated of protein were2,1protein was up-regulated and1protein was down regulated eventually;the expression of protein which down regulated and then increased were10;included expression of6protein expression up-regulated and4protein expression down regulated finally.