Differentially Expressional Proteomics Analysis Of Male And Female Gametophytes Of Ceratoteris Thalicdtroides

Ceratopteris thalictroides (L.) Brongnis is a herbaceous plant species in the genus Ceratopteris in Ceratopteridaceae, which is a model plant of fern studying development,reproduction and sex differentiation. Environmental factors have important influence on the growth and sex differentiation of Ceratopteris thalictroides (L.) Brongnis, and reseaches on the gametophyte growth and the mechanism of sex differentiation of Ceratopteris thalictroides becomes popular. In this paper, we first induced and isolated female and male gametophyte on physiological level, and then studied differential expressed proteins of female and male gametophyte by using proteomic techniques.,which is useful to reveal the fern gametophyte development and sex differentiation mechanism The main results are as follows:1. In the light intensity 2000lx, temperature 20-25℃, the Ceratopteris thalictroides(L.) Brongn spores grown on MS medium will mature after 30 days, 90% of which differentiate into female gametophyte. Density and light intensity has little influence on sex differentiation of Ceratopteris thalictroides gametophyte on the MS culture.2. In the light intensity 2000LX, temperature 20-25℃, culture density of 800 spores / Petri dish, the Ceratopteris thalictroides (L.) Brongnis spores grow on the knops medium will mature after 30 days, 95% of which tend to differentiate into male gametophyte. Density and light intensity has significant influence on sex differentiation of Ceratopteris thalictroides gametophyte on the knop's culture.3. We studied differential protein expression of male and female gametophyte by using two-dimensional electrophoresis technology and mass spectrometry. The results showed that: 1136±55 protein spots was detected in the female gametophyte, and 1130±65 protein spots was detected in the male gametophyte; 57 protein spots, expression changes of which are more than 1.5 times , were analysed by 2D image analysis software -ImageMaster6.0. The expression level of 18 protein spots were higher in the male gametophyte, and 24 protein spots were higher in the female gametophyte. 8 protein spots are only expressed in the female gametophyte, and 7 protein spots are only expressed in the male gametophyte. By MALDI-TOF-TOF / MS analysis and protein database searching, 33 protein spots differentially expressed were identified, which represented 26 species of protein. These proteins mainly involved in photosynthesis (24%,such as:photosystem II oxygen-evolving complex,Rubisco activating enzyme,ribulose-1,5-bisphosphate carboxylase/oxygenase and so on), metabolic (18%,Cystathionine beta-synthase,glyoxalase,Arginase and so on), protein synthesis (6%,translation initiation factor and elongation factor ), protein folding and transport (28%,such as:HSP70,HSP90,molecular chaperone GroEL and so on), energy metabolism (6% , cytosolic malate dehydrogenase and phosphoglycerate mutase), cell structure (12%,centrin,actin 2,actin and so on) and stress and defense (6% , glutathione thiol transferase and Dehydroascorbate Reductase). These results provide a basis of proteomics for further research on the mechanism of gametophyte sex differentiation in ferns.