|
|
|
| Cloning, Subcellular Localization and Expression Analysis of SmbHLH37 Gene in Salvia miltiorrhiza |
| , , , , , , |
|
|
|
|
Abstract Abstract Basic helix-loop-helix (bHLH) transcription factors play important roles in regulating plant growth-development and abiotic stress response. In order to enrich the research content of bHLH transcription factors and find new bHLH transcription factor in regulating secondary metabolite in Salvia miltiorrhiza, SmbHLH37 was cloned based on gDNA and cDNA. The gene sequence of SmbHLH37 was 1 506 bp with one intron of 27 bp, and 1 479 bp complete CDS (GenBank accession No. KP257470.1) encoding 492 amino acids. BlastP result showed that SmbHLH37 protein had high homology with bHLH3 in Sesamum indicum etc. Amino acid sequence analysis showed that SmbHLH37 contained a HLH domain and motif 7. Molecular evolutionary tree of SmbHLH37 protein and all the bHLH transcription factors in Arabidopsis thaliana indicated that SmbHLH37 protein was the closest relative to AtJAM3 (A. thaliana jasmonate-associated MYC2-like 3). Subcellular localization showed that SmbHLH37 protein was mainly localized in the nucleus. The expression pattern disclosed that the expression level of SmbHLH37 was the highest in leaves and the lowest in flowers. qRT-PCR analysis indicated that expression of SmbHLH37 could increase in a short time with 0.1 mmol/L abscisic acid (ABA) and wounding treatment. SmMYC2 and SmbHLH37 expression levels in SmMYC2 overexpressing transgenic S. miltiorrhiza lines OE-8 and OE-12 were detected, and the results showed SmMYC2 expression levels in OE-8 and OE-12 significantly or extremely significantly increased (P＜0.05 or P＜0.001). Meanwhile, the expression levels of SmbHLH37 in OE-8 and OE-12 were similar with SmMYC2 expression. The present data has laid a foundation for studying the function of SmbHLH37 in the future．
|
|
Received: 20 January 2017
Published: 01 June 2017
|
|
|
|
| [1]刘海燕, 冯冬茹, 刘兵, 等.农杆菌介导的蛋白在洋葱表皮细胞的定位研究[J].热带亚热带植物学报, 2009, 17(3):218-222
[2]王庆浩, 陈爱华, 张伯礼.丹参:一种中药研究的模式生物[J].中医药学报, 2009, 37(4):1-3
[3]周宏骏, 武玉翠, 晋鑫鑫, 等.丹参转录因子的克隆及表达模式分析[J].中草药, 2014, 45(23):3449-3455
[4]沈乾, 陆续, 张凌, 等.植物中转录因子功能研究进展[J].上海交通大学学报农业科学版, 2012, 30(6):51-57
[5]Abe H, Urao T, Ito T, et al.Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling[J].Plant Cell, 2003, 15(1):63-78
[6]Castillon, Alicia, Shen, et al.Phytochrome Interacting Factors: central players in phytochrome-mediated light signaling networks[J].Trends in Plant Science, 2007, 12(11):514-521
[7]Dombrecht B, Xue G P, Sprague S J, et al.MYC2 differentially modulates diverse jasmonate-dependent functions in Arabidopsis[J].Plant Cell, 2007, 19(7):2225-2245
[8]Feyissa D N, L?vdal T, Olsen K M, et al.The endogenous GL3,but not EGL3,gene is necessary for anthocyanin accumulation as induced by nitrogen depletion in Arabidopsis rosette stage leaves[J].Planta, 2009, 230(4):747-754
[9]Hua S, Fan H J, Ling H Q.Genome-wide identification and characterization of the bHLH,gene family in tomato[J].BMC Genomics, 2015, 16(1):9-9
[10]Heim M A, Jakoby M, Werber M, et al.The basic helix-loop-helix transcription factor family in plants: a genome-wide study of protein structure and functional diversity[J].Molecular Biology & Evolution, 2003, 20(5):735-747
[11]Li Y G, Song L, Liu M, et al.Advancement in analysis of Salviae miltiorrhizae Radix et Rhizoma (Danshen)[J].Journal of Chromatography A, 2009, 1216(11):1941-1953
[12]Li H, Sun J, Xu Y, et al.The bHLH-type transcription factor AtAIB positively regulates ABA response in Arabidopsis[J].Plant Molecular Biology, 2007, 65(5):655-665
[13]Pires N, Dolan L.Origin and Diversification of Basic-Helix-Loop-Helix Proteins in Plants[J].Molecular Biology & Evolution, 2010, 27(27):862-874
[14]Park K I, Hoshino A.A WD40-repeat protein controls proanthocyanidin and phytomelanin pigmentation in the seed coats of the Japanese morning glory[J].Journal of Plant Physiology, 2012, 169(5):523-8
[15]Qi T, Wang J, Huang H, et al.Regulation of Jasmonate-Induced Leaf Senescence by Antagonism between bHLH Subgroup IIIe and IIId Factors in Arabidopsis[J].Plant Cell, 2015, 27(6):1634-49
[16]Qi T, Huang H, Song S, et al.Regulation of Jasmonate-Mediated Stamen Development and Seed Production by a bHLH-MYB Complex in Arabidopsis[J].The Plant cell, 2015, 27(6):1620-1633
[17]Sasakisekimoto Y, Jikumaru Y, Obayashi T, et al.Basic Helix-Loop-Helix Transcription Factors JASMONATE-ASSOCIATED MYC2-LIKE1 (JAM1),JAM2,and JAM3 Are Negative Regulators of Jasmonate Responses in Arabidopsis[J].Plant Physiology, 2013, 163(1):291-304
[18]Wang X, Morrisnatschke S L, Lee K H.New developments in the chemistry and biology of the bioactive constituents of Tanshen[J].Medicinal Research Reviews, 2007, 27(1):133-148
[19]Wang D, Yao W, Song Y, et al.Molecular characterization and expression of three galactinol synthase genes that confer stress tolerance in Salvia miltiorrhiza[J].Journal of Plant Physiology, 2012, 169(18):1838-1848
[20]Xiong Y, Liu T, Tian C, et al.Transcription Factors in Rice: A Genome-wide Comparative Analysis between Monocots and Eudicots[J].Plant Molecular Biology, 2005, 59(1):191-203
[21]Zhou L, Zuo Z, Chow M S S.Danshen: An overview of its chemistry,pharmacology,pharmacokinetics,and clinical use[J].Journal of Clinical Pharmacology, 2006, 45(12):1345-1359
[22]Zhang X, Luo H, Xu Z, et al.Genome-wide characterisation and analysis of bHLH transcription factors related to tanshinone biosynthesis in Salvia miltiorrhiza[J]. Scientific Reports, 2015, 5:11244.[J].Scientific Reports, 2015, 5(11244):11244-11244
[23]Zhou Y, Sun W, Chen J, et al.SmMYC2a and SmMYC2b played similar but irreplaceable roles in regulating the biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza[J]. Scientific Reports, 2016, 6:22852.[J].Scientific Reports, 2016, 6(22852):22852-22852
|
|
|
|
