Identification And Analysis Of Proteins Related To The Tapping Panel Dryness(TPD) In Hevea Brasiliensis

The Tapping panel dryness (TPD) in Hevea brasiliensis is a complex physiological syndrome. It is one of the important factors of limiting the yield of the natural rubber. In this study, the comparative proteomics method was adopted. The rubber tree clone reyan7-33-97was used as the experimental materials. The differentially expressed proteins from latex C-serums, lutoids and rubber particles in healthy and TPD trees were separated and identified by the two-dimensional gel electrophoresis (2-DE) and mass spectrometry. Bioinformatics was used to analyze the functions of these proteins, and then screen the proteins which were closely related with TPD. The main research results were as follows:1. The identification of TPD related proteins from the C-serums:1168±26and1170±50protein spots were respectively detected in the C-serum protein profiles in healthy and TPD trees. After the mass spectrometry analysis,10spots were successfully identified. These proteins mainly included:the Rub1, AIR9protein, flagellar inner dynein arm heavy chain11, beta-1,3-glucanase, and so on.2. The identification of TPD related proteins from the lutoids:1188±38and1121±45protein spots were detected in the lutoid protein profiles in healthy and TPD trees. After the mass spectrometry analysis,11spots were successfully identified. These proteins were mainly included:chloroplast23kDa polypeptide of photosystem Ⅱ, ATP synthase beta chain, NADP-dependent sorbitol-6-phosphate dehydrogenase, vitamin-b12independent methionine synthase, and so on.3. The identification of TPD related proteins from the rubber particles:498±31and427±19protein spots were detected in the rubber particle protein profiles in healthy and TPD trees. After the mass spectrometry analysis,11spots were successfully identified. These proteins mainly included:REF, enolase1, GRX, GR, actin, HSP, TCTP, farnesyl-diphosphate synthase, ATP synthase subunit alpha, latex-abundant protein, and so on.A total of103differentially expressed proteins were separated from latex C-serums, lutoids and rubber particles. Of them,34proteins were successfully identified, including30down-regulated proteins and4up-regulated ones in TPD trees. Functional classification showed that these proteins were mainly relevant with stress, reactive oxygen metabolism and rubber biosynthesis. This implied that the reactive oxygen metabolism and rubber biosynthesis pathways existed in the process of TPD. Some of these proteins may play key roles in TPD, such as beta-1,3-glucanases GRX, GR, TCTP, HSP, and so on. TCTP and HSP were related with cell apoptosis. The real time quantitative PCR was used to study the differential expression of HbTCTP and HbHSP70genes in healthy and TPD trees. It was found that their expression trends were consistent with ones obtained at the protein level. This research would provide a theoretical basis for clarifying TPD mechanism and the prevention and control of TPD by means of biotechnology.