|
|
Study on Construction of Expression Vector of BoTCTP in Cabbage (Brassica oleracea) and Transferring to Chinese Kale (Brassica alboglabra) |
WANG Jing-Jing1,2,CAO Bi-Hao1,CHEN Guo-Ju1,CHEN Chang-Ming1,CHEN Qing-Hua1,LEI Jian-Jun1* |
1 College of Horticulture, South China Agriculture University, Guangzhou 510642, China; 2 Dezhou Academy of Agricultural Sciences, Dezhou 253015, China |
|
|
Abstract Chinese kale (Brassica alboglabra) is one of the important vegetables in the Cruciferous family. It is often subjected to various adverse effects during the cultivation process, causing great losses to the production. In order to improve the resistance of Chinese kale to stress, the specific primers were designed according to BoTCTP (translationally controlled tumor protein cloned from Brassica oleracea) in this research. The interest fragment was amplified by PCR, and then the overexpression vector was constructed and transferred into Chinese kale by Agrobacterium tumefaciens mediated. The transformants were confirmed by molecular methods and biological assay. Four transgenic plants were obtained, and then taken to design the experiments associated with growth and different stress, such as high temperature, low temperature, salt, heavy metal and so on. The results showed that the transgenic plants grew faster, stronger and had bigger leaves than non-transgenic plants. In the different stress condition, the transgenic plants had obviously stronger resistance than the control, which showed that the BoTCTP was closely related to the growth and had a regulatory effect on the stress resistance of Chinese kale. These results provide some scientific basis for the future research on the stress resistance of Chinese kale.
|
|
|
|
|
|
[1] 曹必好, 雷建军, 陈国菊, 等. 2006. 结球甘蓝转录调控肿瘤蛋白基因(TCTP)的分离与表达特性初步分析[J]. 农业生物技术学报, 14(6): 996-997. (Cao B H, Lei J J, Chen G J, et al. 2006. Isolation and expression analysis of the gene encoding translationally controlled tumor protein (TCTP) in Cabbage[J]. Journal of Agricultural Biotechnology, 14(6): 996-997.)
[2] 曹必好, 雷建军, 宋洪元, 等. 2002. 结球甘蓝cDNA文库的构建和鉴定[J]. 园艺学报, 29(6): 579-580. (Cao B H, Lei J J, Song H Y, et al. 2002. cDNA library construction and identification of cabbage[J]. Acta Horticulturae Sinica, 29(6):579-580.)
[3] 陈长明, 赵祥明, 雷建军, 等. 2016. 基因枪法和农杆菌介导的Bt抗虫基因转化芥蓝[J]. 中国蔬菜, (8): 21-28. (Chen C M, Zhao X M, Lei J J, et al. 2016. Pyramiding of Bt gene into Brassica alboglabra L. H. Bailey by biolistic and Agrobacterium-mediated transformation methods[J]. China Vegetables, (8): 21-28.)
[4] 陈科, 程汉华, 周荣家. 2012. 翻译调节肿瘤蛋白(TCTP)与细胞生长调控[J]. 农业生物技术学报, 20(1): 1-8. (Chen K, Cheng H H, Zhou R J. 2012. Translationally controlled tumour protein (TCTP) and growth regulation of cells[J]. Journal of Agricultural Biotechnology, 20(1): 1-8.)
[5] 丁岳炼, 赵鸿杰, 柯欢, 等. 2014. 山茶花翻译控制肿瘤蛋白CjTCTP基因的电子克隆及生物信息学分析[J]. 黑龙江农业科学, (8): 18-22. (Ding Y L, Zhao H J, Ke H, et al. 2014. Electronic cloning and bioinformatics analysis of CjTCTP gene from Camellia japonica L.[J]. Heilongjiang Agricultural Sciences, (8): 18-22.)
[6] 黄科, 余小林, 吴秋云, 等. 2004. 芥蓝植株再生体系的优化[J].细胞生物学杂志, 26(3): 313-316. (Huang K, Yu X L, Wu Q Y, et al. 2004. The optimization of plant regeneration protocol of Chinese broccoli (Brassica oleracea L. var. alboglabra)[J]. Chinese Journal of Cell Biology, 26(3): 313-316.)
[7] 黄科, 叶纨芝, 余小林, 等. 2007. 农杆菌介导的芥蓝遗传转化体系的建立[J]. 细胞生物学杂志, 29(1): 147-152. (Huang K, Ye W Z, Yu X L, et al. 2007. The establishment of Agrobacterium-mediated transformation system in Brassica oleracea L. var. alboglabra[J]. Chinese Journal of Cell Biology, 29(1): 147-152.)
[8] 雷朋, 何庆玲, 贺芳, 等. 2015. 桑树翻译调控肿瘤蛋白(TCTP)基因的克隆及其在温度胁迫下的表达变化[J]. 蚕业科学, 41(3): 405-409. (Lei P, He Q L, He F, et al. 2015. Cloning of mulberry translationally controlled tumor protein (TCTP) gene and expressional variation under temperature stress[J]. Science of Sericulture, 41(3): 405-409.)
[9] 梁小莲, 李辉亮, 彭世清. 2009. 巴西橡胶树HbTCTP基因的克隆及表达[J]. 分子植物育种, 7(1):188-193. (Liang X L, Li H L, Peng S Q. 2009. Cloning and expression of HbTCTP from Hevea brasiliensis[J]. Molecular Plant Breeding, 7(1): 188-193.)
[10] 林莎, 高帆, 罗洪, 等. 2008. 麻疯树Jc-Tctp1基因的同源性分析及时空表达模式鉴定[J]. 中国生物化学与分子生物学报, 24(8): 727-734. (Lin S, Gao F, Luo H, et al. 2008. Sequencing and expression of translationally controlled tumor protein (TCTP) of Jatropha curcas L.[J]. Chinese Journal of Biochemistry and Molecular Biology, 24(8): 727-734.)
[11] 吕素芳, 郭广君, 蔡永萍. 2006. 翻译控制肿瘤蛋白(TCTP)研究进展[J]. 科学技术与工程, 6(4):424-428. (Lv S F, Guo G J, Cai Y P. 2006. Progress in TCTP[J]. Science Technology and Engineering, 6(4): 424-428.)
[12] 吕亚清. 2009. 芥蓝TCTP基因片段RNAi载体的构建及遗传转化[D]. 硕士学位论文, 华南农业大学, 导师: 雷建军, pp. 30-33. (Lv Y Q. 2009. Construction and genetic transformation of Brassica TCTP gene fragment RNAi vector[D]. Thesis for M.S., South China Agricultural University, Supervisor: Lei J J, pp. 30-33.)
[13] 麻楠, 乔金柱, 孙天杰, 等. 2018. 小麦翻译控制肿瘤蛋白(TCTP)与索马甜类蛋白(TLP)的相互作用[J]. 农业生物技术学报, 26(6): 911-919. (Ma N, Qiao J Z, Sun T J, et al. 2018. Interaction between translationally controlled tumer protein (TCTP) and thaumatin-like protein (TLP) in Wheat (Triticum aestivum)[J]. Journal of Agricultural Biotechnology, 26(6): 911-919.)
[14] 司雨, 陈国菊, 雷建军, 等. 2009. 不同基因型芥蓝硫代葡萄糖苷组分与含量分析[J]. 中国蔬菜, (6): 7-13. (Si Y, Chen G J, Lei J J, et al. 2009. Analysis on composition and content of glucosinolates in different genotypes of Chinese kale[J]. China Vegetables, (6): 7-13.)
[15] 孙勃, 方莉, 刘娜, 等. 2011. 芥蓝不同器官主要营养成分分析[J]. 园艺学报, 38(3): 541-548. (Sun B, Fang L, Liu N, et al. 2011. Studies on main nutritional components of Chinese kale among different organs[J]. Acta Horticulturae Sinica, 38(3): 541-548.)
[16] 王步勇, 李丹蕾, 王峰, 等. 2016. 水稻干尖线虫翻译控制肿瘤蛋白基因的克隆及表达分析[J]. 湖南农业大学学报(自然科学版), 42(1): 20-26. (Wang B Y, Li D L, Wang F, et al. 2016. Clone and expression analysis on the gene of translationally controlled tumor protein from Aphelenchoides besseyi[J]. Journal of Hunan Agricultural University, Natural Sciences, 42(1): 20-26.)
[17] 杨云英, 杨暹. 2002. 温度对芥蓝幼苗叶片内源多胺含量的影响[J]. 华南农业大学学报(自然科学版), 23(3): 9-12. (Yang Y Y, Yang X. 2002. Effect of temperature on endogenous polyamines content of leaves in Chinese kale (Brassica alboglabra Bailey) Seedlings[J]. Journal of South China Agricultural University (Natural Science Edition), 23(3): 9-12.)
[18] 杨云英, 杨暹. 2008. 温度对芥蓝幼苗膜脂过氧化系统的影响[J]. 西南大学学报(自然科学版), 30(8): 124-127. (Yang Y Y, Yang X. 2008. Effect of temperature on membrane lipid peroxidation system in leaves of Chinese kale (Brassica alboglabra Bailey)[J]. Journal of Southwest University (Natural Science Edition), 30(8): 124-127.)
[19] 臧宁, 张美彦, 翟红, 等. 2008. 根癌农杆菌介导的抗除草剂转基因甘薯植株的获得[J]. 农业生物技术学报, 16(1): 103-107. (Zang N, Zhang M Y, Zhai H, et al. 2008. Generation of transgenic herbicide-resistant sweetpotato plants by Agrobacterium tumefaciens-mediated transformation[J]. Journal of Agricultural Biotechnology, 16(1): 103-107.)
[20] 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(4): 555-565.
[21] Berkowitz O, Jost R, Pollmann S, et al. 2008. Characterization of TCTP, the translationally controlled tumor protein, from Arabidopsis thaliana[J]. Plant Cell, 20(12): 3430-3447.
[22] Bommer U A, Thiele B J. 2004. The translationally controlledtumour protein (TCTP)[J]. The International Journal of Biochemistry and Cell Biology, 36(3): 379-385.
[23] Brioudes F, Thierry A M, Chambrier P, et al. 2010. Translationally controlled tumor protein is a conserved mitotic growth integrator in animals and plants[J]. Proceedings of the National Academy of Sciences, 107(37): 16384-16389.
[24] Bruckner F P, Xavier A D, Cascardo R D, 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(5): 672-683.
[25] Cao B H, Lu Y Q, Chen G J, et al. 2010. Functional characterization of the translationally controlled tumor protein(TCTP) gene associated with growth and defense response in cabbage[J]. Plant Cell, Tissue and Organ Culture, 103(2): 217-226.
[26] Chen R Y, Liu H C, Huang Q, et al. 2008. Changes of chlorophyll and anthocyanin content and related enzyme activities of flower stalk in Chinese kale under different light intensities[J]. Acta Horticulture, 769: 103-111.
[27] Chou M X, Xia C C, Feng Z, et al. 2016. A translationally controlled tumor protein gene Rpf41 is required for the nodulation of Robinia pseudoacacia[J]. Plant Molecular Biology, 90(4): 389-402.
[28] Deng M D, Qian H M, Chen L L, et al. 2017. Influence of pre-harvest red light irradiation on main phytochemicals and antioxidant activity of Chinese kale sprouts[J]. Food Chemistry, 222: 1-5.
[29] Deng Z, Chen J, Leclercg J, et al. 2016. Expression profiles, characterization and function of HbTCTP in rubber tree (Hevea brasiliensis)[J]. Frontiers in plant science, 7: 789.
[30] Du J H, Yang P, Kong F H, et al. 2017. Aberrant expression of translationally controlled tumor protein (TCTP) can lead to radioactive susceptibility and chemosensitivity in lung cancer cells[J]. Oncotarget, 8(60): 101922-101935.
[31] Ermolayev V, Weschke W, Manteuffel R. 2003. Comparison of Al-induced gene expression in sensitive and tolerant soybean cultivars[J]. Journal of Experimental Botany, 54(393): 2745-2756.
[32] Hoepflinger M C, Reitsamer J, Geretschlaeger A M, et al. 2013. The effect of translationally controlled tumour protein (TCTP) on programmed cell death in plants[J]. BMC Plant Biology. 13(1): 135.
[33] Hsu Y C, Chern J J, Cai Y, et al. 2007. Drosophila TCTP is essential for growth and proliferation through regulation of dRheb GTPase[J]. Nature, 445(7129): 785-788.
[34] Issarakraisila M, Ma Q F, Turner D W. 2007. Photosynthetic and growth responses of juvenile Chinese kale(Brassica oleracea var. alboglabra) and Caisin (Brassica rapa subsp parachinensis) to water logging and water deficit[J]. Scientia Horticulture, 111(2): 107-113.
[35] Jeon H J, Cui X S, Guo J, et al. 2017. TCTP regulates spindle assembly during postovulatory aging and prevents deterioration in mouse oocyte quality[J]. Elsevier Journal, 1328-1334.
[36] Kim Y M, Han Y J, Hwang O J, et al. 2012. Overexpression of Arabidopsis translationally controlled tumor protein gene AtTCTP enhances drought tolerance with rapid ABA-induced stomatal closure[J]. Molecules and Cells, 33(6):617-626.
[37] Li S, Chen M H, Xiong Q, et al. 2016. Characterization of the translationally controlled tumor protein (TCTP) interactome reveals novel binding partners in human cancer cells[J]. Journal of proteome research, 15(10): 3741-3751.
[38] Lopez A P, Franco A R. 2006. Cloning and expression of cDNA encoding translationally controlled tumor protein from strawberry fruits[J]. Biologia Plantarum, 50(3): 447-449.
[39] Lucibello M, Gambacurta A, Zonfrillo M. 2011. TCTP is a critical survival factor that protects cancer cells from oxidative stress induced cell-death[J]. Experimental Cell Research, 317(17): 2479-2489.
[40] Márcio de C, Márcio L A, Alessandra V N L, et al. 2017. Impacts of the overexpression of a tomato translationally controlled tumor protein (TCTP) in tobacco revealed by phenotypic and transcriptomic analysis[J]. Plant Cell Reports, 36(6): 887-900.
[41] Meng X, Chen Q, Fan H, et al. 2017. Molecular characterization, expression analysis and heterologous expression of two translationally controlled tumor protein genes from Cucumis sativus[J]. PLoS One, 12(9): e0184872
[42] Meng X N, Yu Y , Zhao J Y, et al. 2018. The two translationally controlled tumor protein genes, CsTCTP1 and CsTCTP2, are negative modulators in the Cucumis sativus defense response to sphaerotheca fuliginea[J]. In Plant Science, 9: 544.
[43] Qian H M,Sun B,Miao H Y,et al. 2015. Variation of glucosinolates and quinone reductase activity among different varieties of Chinese kale and improvement of glucoraphanin by metabolic engineering[J]. Food Chemistry, 168: 321-326.
[44] Santa Brigida A B, Dos Reis S P, Costa C N M, et al. 2014. Molecular cloning and characterization of a cassava translationally controlled tumor protein gene potentially related to salt stress response[J]. Molecular Biology Reports, 41(3): 1787-1797.
[45] Sturzenbaum S R, Kille P, Morgan A J. 1998. Identification of heavy metal induced changes in the expression patterns of the translationally controlled tumour protein (TCTP) in the earthworm Lumbricus rubellus[J]. Biochimica et Biophysica Acta, 1398(3): 294-304.
[46] Tsarova K, Yarmola E G, Bubb M R. 2010. Identification of a cofilin-like actin-binding site on translationally controlled tumor protein (TCTP)[J]. FEBS Letters, 584(23): 4756-4760.
[47] Toscano-Morales R, Xoconostle-Cázares B, Martínez-Navarro A C, et al. 2014. Long distance movement of an arabidopsis translationally controlled tumor protein (AtTCTP2) mRNA and protein in tobacco[J]. Frontiers in Plant Science, (5): 705.
[48] Toscano-Morales R, Xoconostle-Cázares B, Cabrera-Ponce J L, et al. 2015. AtTCTP2, an Arabidopsis thaliana homolog of translationally controlled tumor protein, enhances in vitro plant regeneration[J]. Frontiers in Plant Science, (6): 468.
[49] Toscano-Morales R, Xoconostle-Ca′zares B, Mart?′nez-Navarro A C, et al. 2016. AtTCTP2 mRNA and protein movement correlates with formation of adventitious roots in tobacco[J]. Plant Signaling & Behavior,11(3): e1071003.
[50] Tuynder M, Susini L, Prieur S, et al. 2002. Biological models and genes of tumor reversion: Cellular reprogramming through tpt1/TCTP and SIAH-1[J]. Proceedings of the National Academy of Sciences, 99(23): 14976-14981.
[51] Wang L L, Tang Y F, Zhao M J, et al.. 2018. Knockdown of translationally controlled tumor protein inhibits growth, migration and invasion of lung cancer cells[J]. Life Sciences, 193: 292-299.
[52] Wang S, Zhao X F, Wang J X. 2009. Molecular cloning and characterization of the translationally controlled tumor protein from Fenneropenaeus Chinensis[J]. Molecular Biology Reports, 36(7): 1683-1693.
[53] Wang Z Q, Li G Z, Gong Q Q, et al. 2015. OsTCTP, encoding a translationally controlled tumor protein, plays an important role in mercury tolerance in rice[J]. BMC Plant Biology, 15(1): 1-15.
[54] Yenofsky R, cerehini S, kraowczynska A, et al. 1983. Regulation of mRNA utilization in mouse erythroleukemia cells induced to differentiate by exposure to dimethyl sulfoxide[J]. Molecular Cell Bioogy, 3(7):1197-1203.
[55] Zhang J M, Ma L F, Li W, et al. 2014. Cotton TCTP1 gene encoding a translationally controlled tumor protein participates in plant response and tolerance to aphids[J]. Plant Cell Tissue and Organ Culture, 117 (2): 145-156
[56] Zhu Z S, Sun B M, Xu X X, et al. 2016. Overexpression of AtEDT1/HDG11 in Chinese kale (Brassica oleracea var. alboglabra) enhances drought and osmotic stress tolerance[J]. Frontiers in Plant Science, (7): 1285 |
|
|
|