Abstract:BBX (B-box) protein is a subfamily of zinc finger transcription factor protein family, which is widely involved in plant growth and development. And, they also play an important role in abiotic stress response. Eucalyptus is an economic tree species in southern China. However, most cultivated Eucalyptus species are sensitive to abiotic stresses, which made the extension of Eucalyptus cultivation range and the improvement of cultivation benefit limited. In order to enrich gene resources of abiotic stress resistance and provide the basis for molecular assisted breeding for Eucalyptus, total of 21 EgrBBX gene family members were identified from the Eucalyptus grandis genome. Chromosomes distribution, protein sequence structure, and the classes and distribution of cis-elements in promoters were characterized with software of mg2c, Protparam, MEGA and PlantCARE. And, tissue specific expression of EgrBBX family genes and their expression profiling under low temperature, drought and salinity were also analyzed with qRT-PCR. The results showed that EgrBBX genes, named EgrBBX1~21, were distributed at 9 chromosomes. The encoded protein sequences contained classic B1, B2 and CCT domains and could be classified into 5 classes: B1+B2+CCT type ⅠandⅡ, B1+B2, B1+CCT and B1. The results of promoter element analysis indicated that a large number of light-responsive elements and abiotic stress response elements such as ABRE (ABA-responsive element), MBS (MYB binding site), LTR (low-temperature responsiveness), and HSE (heat-stress responsive element) existed in promoter sequences of EgrBBX family gene. qRT-PCR results showed that EgrBBX1, EgrBBX7, EgrBBX9, EgrBBX13 and EgrBBX18 were mainly expressed in leaves, and EgrBBX3, EgrBBX6 and EgrBBX15 had relatively high expression levels in stems. Under the treatments of different time at 4 °C, drought and high- salinity (200 mmol/L NaCl), most EgrBBX gene expression were changed. During the time of low temperature treatment, expression of EgrBBX4, EgrBBX7, EgrBBX8, EgrBBX14, EgrBBX16 and EgrBBX17 showed strong inhibition. EgrBBX8, EgrBBX11, EgrBBX12 and EgrBBX18 showed significant up-regulated expression at 1 d after drought treatment, and the induction effect weakened or disappeared after 2 d. Under high salinity treatment, 13 of 21 EgrBBX genes reached expression peak after 12 h treatment and then gradually decreased. The results in this study provided a basis for further revealing the functions of EgrBBX family in abiotic stress response, and could facilitate the screening of EgrBBX genes which are stress-resistant in Eucalyptus.
杨宁, 从青, 王晓荣, 倪晓祥, 程龙军. 巨桉EgrBBX基因家族鉴定及其在非生物逆境处理下的表达分析[J]. 农业生物技术学报, 2020, 28(4): 658-671.
YANG Ning, CONG Qing, WANG Xiao-Rong, NI Xiao-Xiang, CHENG Long-Jun. Identification of EgrBBX Gene Family and Its Expression Analysis Under Abiotic Stress in Eucalyptus grandis. 农业生物技术学报, 2020, 28(4): 658-671.
[1] 王亚红, 刘缙, 王玉国. 2010. 高质量提取银杏种仁总RNA的改良方法[J]. 中国农学通报, 26(15): 48-52. (Wang Y H, Liu J, Wang Y G.2010. An improved method for RNA isolation from seeds of Ginkgo biloba L[J]. Chinese Agricultural Science Bulletin, 26(15): 48-52.) [2] Bai S, Tao R, Tang Y, et al.2019. BBX 16, a B-box protein, positively regulates light-induced anthocyanin accumulation by activating MYB10 in red pear[J]. Plant Biotechnology Journal, 17(10): 1985-1997. [3] Bowler C, Botto J, Deng X W.2013. Photomorphogenesis, B-Box transcription factors, and the legacy of Magnus Holm[J]. The Plant Cell, 25(4): 1192-1195. [4] Chang C S J, Li Y H, Chen L T, et al.2008. LZF1, a HY5-regulated transcriptional factor, functions in Arabidopsis de-etiolation[J]. The Plant Journal, 54(2): 205-219. [5] Chu Z, 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. [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] Crocco C D, Holm M, Yanovsky M J, et al.2011. Function of B-BOX under shade[J]. Plant Signaling & Behavior, 6(1): 101-104. [8] Crocco C D, Locascio A, Escudero C M, et al.2015. The transcriptional regulator BBX24 impairs DELLA activity to promote shade avoidance in Arabidopsis thaliana[J]. Nature Communications, 6(6): 6202-6209. [9] Crocco C D, Ocampo G G, Ploschuk E L, et al.2018. Heterologous expression of AtBBX21 enhances the rate of photosynthesis and alleviates photoinhibition in Solanum tuberosum[J]. Plant Physiology, 177(1): 369-380. [10] Datta S, Hettiarachchi C, Johansson H, et al.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. [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] Imtiaz M, Yang Y, Liu R, et al.2015. Identification and functional characterization of the BBX24 promoter and gene from chrysanthemum in Arabidopsis[J]. Plant Molecular Biology, 89(1-2): 1-19. [14] Khanna R, Kronmiller B, Maszle D R, et al.2009. The Arabidopsis B-box zinc finger family[J]. The Plant Cell, 21(11): 3416-3420. [15] Kiełbowicz-Matuk A, Rey P, Rorat T.2014. Interplay between circadian rhythm, time of the day and osmotic stress constraints in the regulation of the expression of a Solanum double B-box gene[J]. Annals of Botany, 113(5): 831-842. [16] Klug A, Schwabe J.1995. Protein motifs 5. Zinc fingers[J]. The FASEB Journal, 9(8): 597-604. [17] Laubinger S, Marchal V, Gentilhomme J, 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. [18] Liu J, Shen J, Xu Y, et al.2016. Ghd2, a CONSTANS-like gene, confers drought sensitivity through regulation of senescence in rice[J]. Journal of Experimental Botany, 67(19): 5785-5798. [19] Liu X, Li R, Dai Y, et al.2018. Genome-wide identification and expression analysis of the B-box gene family in the Apple (Malus domestica Borkh.) genome[J]. Molecular Genetics and Genomics, 293(2): 303-315. [20] Liu X, Li R, Dai Y, et al.2019a. A B-box zinc finger protein, MdBBX10, enhanced salt and drought stresses tolerance in Arabidopsis[J]. Plant Molecular Biology, 99: 437-447. [21] Liu Y, Chen H, Ping Q, et al.2019b. The heterologous expression of CmBBX22 delays leaf senescence and improves drought tolerance in Arabidopsis[J]. Plant Cell Reports, 38: 15-24. [22] Meroni G, Diez-Roux G.2005. TRIM/RBCC, a novel class of 'single protein RING finger' E3 ubiquitin ligases[J]. Bioessays, 27(11): 1147-1157. [23] Putterill J, Robson F, Lee K, et al.1995. The CONSTANS gene of Arabidopsis promotes flowering and encodes a protein showing similarities to zinc finger transcription factors[J]. Cell, 80(6): 847-857. [24] Sarmiento F.2013. The BBX subfamily IV: Additional cogs and sprockets to fine-tune light-dependent development[J]. Plant Signaling & Behavior, 8(4): e23831. [25] Seo E, Kim S, Yeom S I, et al.2016. Genome-wide comparative analyses reveal the dynamic evolution of nucleotide-binding leucine-rich repeat gene family among Solanaceae plants[J]. Frontiers in plant science, 7(1205): 1-13. [26] Takatsuji H.1998. Zinc-finger transcription factors in plants[J]. Cellular and Molecular Life Sciences CMLS, 54(6): 582-596. [27] Tiwari S B, Shen Y, Chang H C, et al.2010. The flowering time regulator CONSTANS is recruited to the FLOWERING LOCUS T promoter via a unique cis-element[J]. New Phytologist, 187(1): 57-66. [28] Vaishak K, Yadukrishnan P, Bakshi S, et al.2019. The B-box bridge between light and hormones in plants[J]. Journal of Photochemistry and Photobiology B: Biology, 191(2019): 164-174. [29] 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. [30] Yano M, Katayose Y, Ashikari M, et al.2000. Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS[J]. The Plant Cell, 12(12): 2473-2483. [31] Zou Z, Wang R, Wang R, et al.2017. Genome-wide identification, phylogenetic analysis, and expression profiling of the BBX family genes in pear[J]. The Journal of Horticultural Science and Biotechnology, 93(1): 37-50.