Thioredoxins play critical roles in the maintenance of cellular redox homeostasis by regulating the structure and function of the target proteins through catalyzing thiol-disulfide exchange reactions. Previous research has confirmed that the y-type thioredoxin HbTRXy2 of rubber tree (Hevea brasiliensis) has a strong antioxidant function. To investigate the molecular mechanism of HbTRXy2 in enhancing antioxidant defense, a yeast two-hybrid library of rubber tree and the bait vector pGBKT7-HbTRXy2 for HbTRXy2 were constructed, and the interacting proteins of HbTRXy2 were identified via yeast two-hybrid in this study. The results indicated that the secondary library constructed in this study had a recombination rate of 100%, an average length of inserted fragments greater than 1000 bp, and a capacity of approximately 3.80×106 CFU/mL. It was identified that the bait vector has no toxicity and self-activating activity in yeast and could be used for yeast two-hybrid screening. After testing, it was found that the bait vector had no toxicity and self-activating activity in yeast and could be used for yeast two-hybrid screening. Using the co-transformation method, 24 proteins interacting with HbTRXy2 were screened from the constructed library. Bioinformatics analysis revealed that the functions of the candidate interacting proteins encompassed redox processes, stress responses, ATP binding and metabolism, Calvin cycle, single-carbon metabolism, carbon fixation, methionine biosynthesis, lipid A biosynthesis, metal ion binding or transport, transmembrane transport, cell volume regulation, phosphate ion homeostasis, protein phosphorylation, protein folding, etc. The results would lay the foundation for further revealing the function and mechanism of HbTRXy2 in rubber tree.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |