|
|
Cloning and Expression of Transcription Factor SpBBX1 in Solanum pennellii and Preliminary Analysis of Its Stress Tolerance |
ZHAO Si-Fang1,2, ZHOU Tao1,2, HU Jia-Hui1,2, LAN Hai-Yan1,*, YU Qing-Hui2,* |
1. Xinjiang Key Laboratory of Biological Resources and Genetic Engineering/College of Life Science and Technology, Xinjiang University, Urumqi 830046, China; 2. Institute of Horticultural Crops, Xinjiang Academy of Agriculture Sciences, Urumqi 830091, China |
|
|
Abstract B-box (BBX) transcription factor plays an important role in the photomorphogenesis of seedlings, regulation of flowering photoperiod, avoidance of light and other growth and developmental processes, as well as in response to the biotic and abiotic stresses. In this study, a BBX protein was obtained from Solanum pennellii and named as SpBBX1 (Gene ID: Sopen02g034260). In order to study its function, the physicochemical properties, protein structure, phylogenetic tree, subcellular localization and prokaryotic expression were analyzed. The results showed that SpBBX1 contained a complete open reading frame of 1 173 bp, encoding 390 amino acids, and was located in the nucleus; phylogenetic tree analysis showed its amino acid sequence was closely related to cultivated tomato protein SlCONSTANS1 (Gene ID: Solyc02g089540) and potato protein StB-box1 (Gene ID: PGSC0003DMP400017796); SDS-PAGE and Western blot results showed that the molecular weight of recombinant SpBBX1 protein was about 48 kD; qRT-PCR analysis found that SpBBX1 was expressed in stem, leaf, and flower, with the highest expression in leaf. The prokaryotic recombinant strain Escherichia coli BL21::pET-30a-SpBBX1 showed significantly lower growth than the control strain in response to NaCl or polyethylene glycol (PEG) stress (P<0.05). These results suggested that SpBBX1 gene may have a negative regulatory effect in response to stress, the data could provide a theoretical basis for further research on plant BBX function.
|
Received: 22 March 2021
|
|
Corresponding Authors:
* lanhaiyan@xju.edu.cn; yuqinghui98@sina.com
|
|
|
|
[1] 田静. 2014. 番茄抗逆相关基因ShZFP1的克隆与功能分析[D]. 硕士毕业论文, 华中农业大学, 导师: 叶志彪. pp. 6-7. (Tian J.2014. Cloning and functional analysis of stress-related gene ShZFP1 in tomato[D]. Thesis fo M.S., Huazhong Agricultural University, Supervisor: Ye Z B, pp. 6-7.) [2] 赵悦. 2019. 中国番茄酱出口价格下滑原因及应对策略[J]. 河北企业, 421(32): 7-12. (Zhao Y.2019. Reasons for falling export price of Chinese ketchup and countermeasures[J]. Hebei Enterprise, 421(32): 7-12.) [3] Chang C S J, Li Y H, Chen L T, et al.2010. LZF1, a HY5-regulated transcriptional factor, functions in Arabidopsis deetiolation[J]. The Plant Journal, 54(2): 205-219. [4] Chen J, Chen J Y, Wang J N, et al.2012. Molecular characterization and expression profiles of MaCOL1, a CONSTANS-like gene in banana fruit[J]. Gene, 496(2): 110-117. [5] Chu Z N, Wang X, Li Y, et al.2016. Genomic organization, phylogenetic and expression analysis of the B-BOX gene family in tomato[J]. Frontiers in Plant Science, 7: 1552-1566. [6] Crocco C D, Botto J F.2013. BBX proteins in green plants: Insights into their evolution, structure, feature and functional diversification[J]. Gene, 531(1): 44-52. [7] Datta S.2006. Arabidopsis constans-like3 is a positive regulator of red light signaling and root growth[J]. The Plant Cell, 18(1): 70-84. [8] Datta S, Hettiarachchi C, Holm J M.2007. Salt tolerance homolog2, a B-Box protein in Arabidopsis that activates transcription and positively regulates light-mediated development[J]. The Plant Cell, 19(10): 3242-3255. [9] Datta S, Johansson H, Hettiarachchi C, et al.2008. lzf1/salt tolerance homolog3, an Arabidopsis B-Box protein involved in light-dependent development and gene expression, undergoes COP1-mediated ubiquitination[J]. The Plant Cell, 20(9): 2324-2338. [10] Fan X Y, Sun Y, Cao D M, et al.2012. BZS1, a B-box protein, promotes photomorphogenesis downstream of both brassinosteroid and light signaling pathways[J]. Molecular Plant, 5(3): 591-600. [11] Gangappa S N, Botto J F.2014. The BBX family of plant transcription factors[J]. Trends in Plant Science, 19(7): 460-470. [12] Gangappa S N, Crocco C D, Johansson H, et al.2013. The Arabidopsis B-box protein BBX25 interacts with HY5, negatively regulating BBX22 expression to suppress seedling photomorphogenesis[J]. The Plant Cell, 25(4): 1243-1257. [13] Gendron J M, Pruneda-Paz J L, Doherty C J, et al.2012. Arabidopsis circadian clock protein, TOC1, is a DNA-binding transcription factor[J]. Proceedings of the National Academy of Sciences of the USA, 109(8): 3167-3172. [14] Griffiths S, Dunford R P, Laurie C D A.2003. Laurie, the evolution of CONSTANS-Like gene families in barley, rice, and Arabidopsis[J]. Plant Physiology, 103(4): 1855-1867 [15] Gao H, Song A, Zhu X, et al.2012. The heterologous expression in Arabidopsis of a chrysanthemum Cys2/His2 zinc finger protein gene confers salinity and drought tolerance[J]. Planta, 235(5): 979-993. [16] Hao L, Shuangyu D, Dayuan S, et al.2016. CONSTANS-Like 9 (OsCOL9) interacts with receptor for activated C-kinase 1 (OsRACK1) to regulate blast resistance through salicylic acid and ethylene signaling pathways[J]. PLOS ONE, 11(11): e0166249. [17] Holtan H E, Bandong S, Marion C M, et al.2011. BBX32, an Arabidopsis B-Box protein, functions in light signaling by suppressing HY5-regulated gene expression and interacting with STH2/BBX21[J]. Plant Physiology, 156(4): 2109-2123. [18] Indorf M, Cordero J, Neuhaus G, et al.2010. Salt tolerance (STO), a stress-related protein, has a major role in light signalling[J]. Plant Journal, 51(4): 563-574. [19] Jang S, Marchal V, Panigrahi K C, et al.2008. Arabidopsis COP1 shapes the temporal pattern of CO accumulation conferring a photoperiodic flowering response[J]. The Embo Journal, 27(8): 1277-1288. [20] Kumagai T, Ito S, Nakamichi N, et al.2008. The common function of a novel subfamily of B-Box zinc finger proteins with reference to circadian-associated events in Arabidopsis thaliana[J]. Bioscience Biotechnology & Biochemistry, 72(6): 1539-1549. [21] Liu X, Li R, Dai Y, et al.2019. A B-box zinc finger protein, MdBBX10, enhanced salt and drought stresses tolerance in Arabidopsis[J]. Plant Molecular Biology, 99(5): 437-447. [22] Ma G P, Zhao D Q, Wang T W, et al.2018. BBX32 interacts with AGL24 involved in flowering time control in Chinese cabbage (Brassica rapa L. ssp. pekinensis)[J]. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 47(1): 34-45. [23] Massiah M A, Matts J A B, Short K M, et al.2007. Solution structure of the MID1 B-box2 CHC(D/C)C(2)H(2) zinc-binding domain: Insights into an evolutionarily conserved RING fold[J]. Journal of Molecular Biology, 369(1): 1-10. [24] Massiah M A, Simmons B N, Short K M, et al.2006. Solution structure of the RBCC/TRIM B-box1 domain of human MID1: B-box with a RING[J]. Journal of Molecular Biology, 358(2): 532-545. [25] Nakazato T, Warren D L, Moyle L C.2010. Ecological and geographic modes of species divergence in wild tomatoes[J]. American Journal of Botany, 97(4): 680-693. [26] Rajnish K, Brent K, Don R, et al.2009. The Arabidopsis B-Box zinc finger family[J]. The Plant Cell, 21(11): 3416-3420. [27] Rajnish K, Yu S, Gabriela T O, et al.2006. Functional profiling reveals that only a small number of phytochrome-regulated early-response genes in Arabidopsis are necessary for optimal deetiolation[J]. The Plant Cell, 18(9): 2157-2171. [28] Reymond A, Meroni G, Fantozzi A, et al.2014. The tripartite motif family identifies cell compartments[J]. The EMBO Journal, 20(9): 2140-2151. [29] Robson F, Costa M M, Hepworth S R, et al.2001. Functional Importance of conserved domains in the flowering-time gene CONSTANS demonstrated by analysis of mutant alleles and transgenic plants[J]. Plant Journal, 28(6): 619-631. [30] Sánchez JP, Duque P, Chua N H.2010. ABA activates ADPR cyclase and cADPR induces a subset of ABA-responsive genes in Arabidopsis[J]. Plant Journal, 38(3): 381-395. [31] Sascha L, Virginie M, José G, et al.2006. Arabidopsis SPA proteins regulate photoperiodic flowering and interact with the floral inducer CONSTANS to regulate its stability[J]. Development, 133(16): 3213-3222. [32] Shuuichi N, Tetsuo T.2003. Salt tolerance-related protein STO binds to a Myb transcription factor homologue and confers salt tolerance in Arabidopsis[J]. Journal of Experimental Botany, 54(391): 2231-2237. [33] Soitamo A J, Piippo M, Allahverdiyeva Y, et al.Light has a specific role in modulating Arabidopsis gene expression at low temperature[J]. Bmc Plant Biology, 2008, 8(1): 13. [34] Takuhara Y, Kobayashi M, Suzuki S.2011. Low-temperature-induced transcription factors in grapevine enhance cold tolerance in transgenic Arabidopsis plants[J]. Journal of Plant Physiology, 168(9): 967-975. [35] Veronica L, Martha S, Charles S.1996. Two classes of plant cDNA clones differentially complement yeast calcineurin mutants and increase salt tolerance of wild-type yeast[J]. Journal of Biological Chemistry, 271(22): 12859-12866. [36] Yan H, Marquardt K, Indorf M, et al.2011. Nuclear localization and interaction with COP1 are required for STO/BBX24 function during photomorphogenesis[J]. Plant Physiology, 156(4): 1772-1782. [37] Wang Q, Tu X, Zhang J, et al.2013. Heat stress-induced BBX18 negatively regulates the thermotolerance in Arabidopsis[J]. Molecular Biology Reports, 40(3): 2679-2688. |
|
|
|