Protemomic Analysis And Physiology Study On Fertility Restoration Of Indica Cms Lines In Rice Under High Temperature

In order to better understand the mechanisms of fertility restoration of CMS lines in indica rice under high temperature.the experiment,which selected Tianfeng A,Wufeng A as the materials and their maintainer line Tianfeng B,Wufeng B as control varieties,was carried out to analyze the physiological and biochemical characteristics of three different tissues(Second leaf,Flag leaf,young panicle)during the stamen and pistil formation stage and pollen filling stage under high or normal temperature.At the same time,proteome analysis was conducted to identify the young panicle proteins in poll mother cells meiosis of three different samples(Tianfeng A performed fertility,Tianfeng A performed sterility,Tianfeng B performed fertility).The main results show as follows:1.Compared with normal temperature(24?)treatment,the proportion of abortive pollen in Tianfeng A and Wufeng A was reduced under high temperature(32 ?),meanwhile,the proportion of stained abortive pollen was increased,and the fertile pollen was appeared and some spikes was selfing.The self seed-setting of Tianfeng A and Wufeng A were 5.40% and 3.17% respectively.But the Tianfeng B and Wufeng B under high temperature treatment turned to be less rate of fertile pollen,more abortive pollen,and less proportion of selfing.2.Some pollen of Tianfeng A and Wufeng A was being fertility restoration under high temperature,and the physiological and biochemical indexes in Tianfeng A and Wufeng A in the young spike differentiation stage were different with those under normal temperature treatment.Under the high temperature treatment,the MDA content in both leaves and young spike of Tianfeng A and Wufeng A were increased;the POD activity in the second leaf and young spike was increased;the soluble protein content in leaves was increased,and the soluble protein content in young spike increased significantly during pollen mother cell formation stage and pollen filling stage,the soluble protein content in young spike was closing to that of maintainer line at pollen filling stage;the proline content was also increased in both leaves and young spike,especially,proline content of young spike was 2 times of Tianfeng A's at pollen filling stage,which was about two thirds of that of the maintainer;the soluble sugar and the starch content of leaves were obviously decreased,but the soluble sugar content of young spike during pollen mother cell formation stage to pollen filling stage was increased.The changes of physiological and biochemical indexes indicate that the cell membrane of rice was influenced under the high temperature treatment,then substance transmembrane transport may be affected,and the redox balance in CMS line altered,soluble protein and proline content of leaves and spikes increased,material and energysupply in young spikes increased,more material and energy could be used to forming pollen mother cells and the filling of pollen content.In short,these changes of physiological and biochemical indexes may be related to pollen fertility restoration.3.Based on the 2-DE analyses,81 spots was showed abundance changes at least 2fold(7)p<0.05(8)among Tianfeng A under high temperature(7)32?(8)treatment,Tianfeng A under normal temperature(7)24 ?(8)treatment and Tianfeng B under normal temperature(7)24?(8),which were identified and quantified in pollen mother cell meiosis young spikes,Compared with Tianfeng A(7)24?(8),46 spots was performed abundance 2 more fold of Tianfeng A(7)32?(8)treatment and Tianfeng B(7)24?(8),which were chosed to be identified by mass spectrometry techniques.30 spots were up-regulated and other 16 spots were down-regulated,And 45 spots were matched successfully.These differentially expressed proteins were mainly involved in proteins response to stimulation,transcription,protein synthesis,protein processing,proteolytic proteins,redox homeostasis,crbohydrate and energy metabolism,signal transduction,lipid metabolism,cellular processing.Among the differentially expressed proteins,acidic ribosomal protein,Methyl-CPG binding domain containing protein,deoxyuridine triphosphatase,disulfide isomerase,proline-rich proteine,nuclear RNA binding protein A,Ribosome recycling factor,S-adenosyl-L-methionine synthetase,and other proteins were probably related to the fertility expression of CMS line of rice.