Abstract：Abstract Translationally controlled tumor protein (TCTP) exists widely in eukaryotic cells. TCTP was involved in a variety of cellular processes including mitotic regulation, DNA damage repair, and plant resistance to pathogen infection. Thaumatin-like protein (TLP) is a pathogenesis related protein in plants and is thought to participate in the various processes of plants in response to pathogens infection. Previous work in this laboratory has demonstrated that TaTCTP was involved in the response of wheat (Triticum aestivum) to Puccinia triticina infection. This study focused on the interaction between TCTP and TLP. Vectors were constructed for yeast two-hybrid and Bi-molecular fluorescence complementation (BiFC), respectively. Through yeast two-hybrid test, it was found that yeast AH109 carried both TCTP and TLP could grow on SD-Leu/-Trp/-His and SD-Leu/-Trp/-His/-Ade medium and have detectable α- galactosidase gene (MEL1) reporter activity. This result indicated that there is a physical interaction between TaTCTP and TaTLP. Through the BiFC test, it was found that a strong yellow fluorescence signal could be observed in tobacco epidermis cytoplasm when co-transforming Agrobacterium with TCTP and TLP. These results indicated that TaTCTP and TaTLP can interact with each other in cytoplasm. This study lay a foundation for further exploring the function and mechanism of TCTP in wheat resistance to Puccinia triticina infection.
白志英, 王冬梅, 候春燕, 等. 2003. 小麦叶锈菌侵染过程的显微和超微结构[J]. 细胞生物学杂志, 25: 393-397. (Bai Z Y, Wang D M, Hou C Y, et al. 2003. Microscopic and ultrastructure of the wheat leaf rust infecting process[J]. Journal of Cell Biology, 25: 393-397.) 白志英, 王冬梅, 陆丙社, 等. 2003. 小麦与叶锈菌互作过程中细胞程序性死亡的细胞学观察[J]. 实验生物学报, 36: 353–360. (Bai Z Y, Wang D M, Lu B S, et al. 2003. Cytological observation of the process of programmed cell death in the interaction between wheat and leaf rust[J]. Journal of Experimental Biology, 36: 353–360.) 冯洁. 2008. 酵母双杂交技术在植物与病原物识别及信号传导研究中的应用[J]. 农业生物技术学报, 16(3):365-372. (Feng J. 2008. Impact of Yeast Two-hybrid Technology on Deciphering Plantpathogen Recognition and Signal Transduction Events[J]. Journal of Agricultural Biotechnology, 2008, 16(3):365-372.) 牛苏燕, 蒋素华, 武思, 等. 2012. 甜蛋白thaumatin基因高效植物表达载体的构建. 江西农业学报, 24: 109–111. (Niu S Y, Jiang S H, Wu S, et al. 2012. Construction of Highly Efficient Plant Expression Vector of Thaumatin Gene[J]. Journal of Jiangxi Agricultural University, 24: 109–111.) 王丰青, 张重义, 童治军,等. 2012. 应用双分子荧光互补(BiFC)方法分析烟草中介体亚基之间的互作[J]. 农业生物技术学报, 20(1):38-47. (Wang F Q, Zhang Z Y, Tong Z J, et al. 2012. Interactions among Mediator Subunits of Tobacco by Bimolecular Fluorescence Complementation (BiFC) Method[J]. Journal of Agricultural Biotechnology, 2012, 20(1):38-47.) 张立峰. 2011. TcLr19与叶锈菌互作早期的酵母双杂交cDNA文库构建及TaTCTP表达分析[D]. 硕士学位论文,河北农业大学,导师:王冬梅. (Zhang L F. 2011. Construction of Yeast Two-hybrid cDNA Library and Expression Analysis of TaTCTP in Early TcLr19 Interaction with Leaf Rust[D]. Master Thesis. Agricultural University of Hebei, Supervisor: Wang D M.) 郑舒扬, 王艳红, 肖莹, 等. 2014. 白粉菌诱导下感病小麦京411中TCTP基因的表达分析[J]. 生物技术通报; 9: 72–77. (Zhen S Y, Wang Y H, Xiao Y, et al. 2014. Analysis of TCTP gene expression in susceptible wheat Jing 411 induced by powdery mildew[J]. Biotechnology Bulletin, 9: 72–77.) Abad LR, D’Urzo MP, Liu D, et al. 1996. Antifungal activity of tobacco osmotin has specificity and involves plasma membrane permeabilization[J]. Plant Science, 118: 11–23. Bruckner FP, Xavier ADS, Cascardo RDS, et al. 2017. Translationally controlled tumour protein (TCTP) from tomato and Nicotiana benthamiana is necessary for successful infection by a potyvirus[J]. Molecular Plant Pathology, 18: 672–683. Berkowitz O, Jost R, Pollmann S, et al. 2008. Characterization of TCTP, the Translationally Controlled Tumor Protein, from Arabidopsis thaliana[J]. Plant Cell Online, 20: 3430–3447. Bommer UA, Thiele BJ. 2004. The translationally controlled tumour protein (TCTP)[J].The International Journal of Biochemistry and Cell Biology, 36: 379–385. Bazile F, Pascal A, Arnal I, et al. 2009. Complex relationship between TCTP, microtubules and actin microfilaments regulates cell shape in normal and cancer cells[J]. Carcinogenesis, 30: 555–565. Cao J, Lv Y, Hou Z, et al. 2016. Expansion and evolution of thaumatin-like protein (TLP) gene family in six plants[J]. Plant Growth Regulation, 79: 299–307. Chen L, Qian J, Qu S, et al. 2008. Identification of specific fragments of HpaG Xooc, a harpin from Xanthomonas oryzae pv. oryzicola, that induce disease resistance and enhance growth in plants[J]. Phytopathology, 98: 781–791. Dhekney SA, Li ZT, Gray DJ. 2011. Grapevines engineered to express cisgenic Vitis vinifera thaumatin-like protein exhibit fungal disease resistance[J]. In Vitro Cellular and Developmental Biology Plant, 47: 458–466. Estrada-Navarrete G, Cruz-Mireles N, Lascano R, et al. 2016. An Autophagy-Related Kinase Is Essential for the Symbiotic Relationship between Phaseolus vulgaris and Both Rhizobia and Arbuscular Mycorrhizal Fungi[J]. Plant Cell, 28: 2326–2341. Fujise K, Li F, Zhang D, et al. 2002. Characterization of Fortilin , a Novel Antiapoptotic Protein Characterization of Fortilin , a Novel Antiapoptotic Protein[J]. Journal of Biological Chemistry, 276: 47542–47549. Gupta M, Yoshioka H, Ohnishi K, et al. 2013. A translationally controlled tumor protein negatively regulates the hypersensitive response in nicotiana benthamiana. Plant Cell Physiology, 54: 1403–1414. Hoepflinger MC, Reitsamer J, Geretschlaeger AM, et al. 2013. The effect of translationally controlled tumour protein (TCTP) on programmed cell death in plants[J]. BMC Plant Biology, 13: 135. Jayasankar S, Li Z, Gray DJ. 2003. Constitutive expression of Vitis vinifera thaumatin-like protein after in vitro selection and its role in anthracnose resistance[J]. Functional Plant Biology, 30: 1105–1115. Kubiak JZ, Bazile F, Pascal A, et al. 2008. Temporal regulation of embryonic M-phases[M]. Folia Histochemica Et Cytochemica , pp. 5–9. Misra RC, Sandeep, Kamthan M, et al. 2016. A thaumatin-like protein of Ocimum basilicum confers tolerance to fungal pathogen and abiotic stress in transgenic Arabidopsis[J]. Scientific Reports, 6: 25340. Meyers BC, Morgante M, Michelmore RW. 2002. TIR-X and TIR-NBS proteins: Two new families related to disease resistance TIR-NBS-LRR proteins encoded in Arabidopsis and other plant genomes[J]. Plant Journal, 32: 77–92. Nielsen H V, Johnsen AH, Sanchez JC, et al. 1998. Identification of a basophil leukocyte interleukin-3-regulated protein that is identical to IgE-dependent histamine-releasing factor[J]. Allergy, 53: 642–652.） Petre B, Major I, Rouhier N, et al. 2011. Genome-wide analysis of eukaryote thaumatin-like proteins (TLPs) with an emphasis on poplar[J]. BMC Plant Biology, 11: 33–48. Steuernagel B, Periyannan SK, Hernández-Pinzón I, et al. 2016. Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture[J]. Nature Biotechnology, 34: 652–655. Sparkes IA, Runions J, Kearns A, et al. 2006. Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants[J]. Nature Protocols, 1: 2019–2025. Song YH, Song NY, Shin SY, et al. 2008. Isolation of CONSTANS as a TGA4/OBF4 interacting protein[J]. Molecules and Cells, 25: 559–565. Stokes TL, Kunkel BN, Richards EJ. 2002. Epigenetic variation in Arabidopsis disease resistance[J]. Genes and Development, 16: 171–182. Tao J, Cao Y, Chen H, et al. 2008. Tobacco TCTP interacts with ethylene receptor NTHK1 and enhances plant growth through promotion of cell proliferation[J]. Plant Physiology, 169: 96–114. Tuynder M, Fiucci G, Prieur S, et al. 2004. Translationally controlled tumor protein is a target of tumor reversion[J]. Proceedings of the National Academy of Sciences, 101: 15364–15369. Wang ZQ, Li GZ, Gong QQ, et al. 2015. OsTCTP, encoding a translationally controlled tumor protein, plays an important role in mercury tolerance in rice[J]. BMC Plant Biology, 15: 123–137. Wang Q, Li F, Zhang X, et al. 2011. Purification and characterization of a CkTLP protein from Cynanchum komarovii seeds that confers antifungal activity[J]. PLoS One, 6: 1–10. Yan X, Qiao H, Zhang X, et al. 2017. Analysis of the grape (Vitis vinifera L.) thaumatin-like protein (TLP) gene family and demonstration that TLP29 contributes to disease resistance[J]. Scientific Reports, 7: 4269. Zhang Z, Chen J, Su Y, et al. 2015. TaLHY, a 1R-MYB transcription factor, plays an important role in disease resistance against stripe rust fungus and ear heading in wheat[J]. PLoS One, 10: 1–13.
