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| Expression Pattern Analysis of Regulatory Genes Responsible for Indolic Glucosinolate Biosynthesis in Arabidopsis thaliana |
| SHENG Yu-Xin, PENG Yi-Fang, ZHU Xing-Yu, JIN Feng, KONG Wen-Wen, LI Jing* |
| College of Life Science, Northeast Agricultural University, Harbin 150030, China |
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Abstract Indolic glucosinolates (IGS) play a crucial role in plant pathogen defense responses, MYB (myeloblastosis) 34, MYB51 and MYB122 are 3 important transcription factors that regulate indolic glucosinolate biosynthesis. This study systematically studied the temporal and spatial expression patterns of 3 transcription factor genes and their responses to hormones, which could provide data for further study on the disease resistance mechanism of indolic glucosinolates. Using Arabidopsis thaliana genomic DNA as a template, the promoter sequences of MYB34, MYB51 and MYB122 genes were amplified, plant expression vectors with GUS (β-glucuronidase coding gene) expression driven by the promoters of MYB34, MYB51, and MYB122, respectively were constructed and transferred into Arabidopsis thaliana. PMYB34::GUS, PMYB51::GUS and PMYB122::GUS transgenic lines were confirmed. Through the detection of GUS activity, the temporal and spatial expression patterns of 3 genes and the response to several hormones were analyzed. Gene expression analysis showed that the temporal and spatial expression patterns of MYB34, MYB51 and MYB122 genes were similar. All the 3 genes generally expressed in various stages of growth and development and in various tissues, in the cotyledons, hypocotyls and radicles of seedlings, and the vascular bundles of roots, stems, leaves, flowers, and pods of mature plants. However, the 3 genes showed their own specific expression pattern. The expression of MYB51 performed higher in ground tissue than in root tissue, while the MYB34 and MYB122 were mainly expressed in the roots. Hormone responsiveness analysis showed that MYB51 was induced by salicylic acid (SA) and MYB34 was induced by methyl jasmonate (MeJA). Compared with MYB34 and MYB51, MYB122 was not sensitive to hormone stimulation. Under the experimental conditions of this study, the expression level of MYB122 did not change significantly after several hormone treatments. The above research results suggested that the metabolism of indolic glucosinolates might be regulated by different signaling pathways in different tissues of the plant. This study preliminarily clarified the temporal and spatial expression characteristics of MYB34, MYB51 and MYB122, and provided basic data for further revealing the regulation mechanism of indolic glucosinolate biosynthesis.
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Received: 02 October 2018
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Corresponding Authors:
lijing@neau.edu.cn
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1 Celenza J L, Normanly J A, Smolen G A.2005. The Arabidopsis ATR1 MYB transcription factor controls indolic glucosinolate homeostasis[J]. Plant Physiology, 137(1): 253-262.
2 Clough S J, Bent A F.1998. Floral dip: A simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J]. Plant Journal, 16(6): 735-743.
3 Fahey J W, Zalcmann A T, Talalay P.2001. The chemical diversity and distribution of glucosinolates and isothiocyanates among plants[J]. Phytochemistry, 56(1): 5-51.
4 Froger A, Hall J E.2007. Transformation of plasmid DNA into E. coli using the heat shock method[J]. Journal of Visualized Experiments, (6): 253.
5 Frerigmann H, Gigolashvili T.2014. MYB34, MYB51, and MYB 122 distinctly regulate indolic glucosinolate biosynthesis in Arabidopsis thaliana[J]. Molecular Plant, 7(5): 814-828.
6 Frerigmann H, Glawischnig E, Gigolashvili T.2015. The role of MYB34, MYB51 and MYB122 in the regulation of camalexin biosynthesis in Arabidopsis thaliana[J]. Front Plant Science, 25(6): 654.
7 Frerigmann H, Piślewska-Bednarek M, Sánchez-Vallet A, et al.2016. Regulation of pathogen-triggered tryptophan metabolism in Arabidopsis thaliana by MYB transcription factors and indole glucosinolate conversion products[J]. Molecular Plant, 9(5): 682-695.
8 Gigolashvili T, Berger B, Mock H P, et al.2007. The transcription factor HIG1/MYB51 regulates indolic glucosinolate biosynthesis in Arabidopsis thaliana[J]. Plant Journal, 50(5): 886-901.
9 Hull A K, Celenza J L.2000. Arabidopsis cytochrome P450s that catalyze the first step of tryptophan-dependent indole-3-acetic acid biosynthesis[J]. Proceedings of the National Academy of Science of the USA, 97(5): 2379-2384.
10 Lee S, Kaminaga Y, Cooper B.2012. Benzoylation and sinapoylation of glucosinolate R-groups in Arabidopsis[J]. Plant Journal, 72(3): 411-422.
11 Megna B W, Carney P R, Nukaya M, et al.2016. Indole-3-carbinol induces tumor cell death: Function follows form[J]. Journal of Surgical Research, 204(1): 47-54.
12 Mikkelsen M D, Hansen C H, Wittstock U.2000. Cytochrome P450 CYP79B2 from Arabidopsis catalyzes the conversion of tryptophan to indole-3-acetaldoxime, a precursor of indole glucosinolates and indole-3-acetic acid[J]. Journal of Biological Chemistry, 275(43): 33712-33717.
13 Nour-Eldin H H, Hansen B G, Norholm M H, et al.2006. Advancing uracil-excision based cloning towards an ideal technique for cloning PCR fragments[J]. Nucleic Acids Research, 34(18): e122.
14 Obayashi T, Hayashi S, Saeki M, et al.2009. ATTED-II provides coexpressed gene networks for Arabidopsis[J]. Nucleic Acids Research, 37: D987-D991.
15 Pfalz M, Mukhaimar M, Perreau F, et al.2016. Methyl Transfer in glucosinolate biosynthesis mediated by indole glucosinolate o-methyltransferase 5[J]. Plant Physiology, 172(4): 2190-2203.
16 Piotrowski M, Schemenewitz A, Lopukhina A, et al.2004. Desulfoglucosinolate sulfotransferases from Arabidopsis thaliana catalyze the final step in the biosynthesis of the glucosinolate core structure[J]. Journal of Biological Chemistry, 279(49): 50717-50725.
17 Seo M S, Kim J S.2017. Understanding of MYB transcription factors involved in glucosinolate biosynthesis in Brassicaceae[J]. Molecules, 22(9): 1549.
18 Stracke R, Werber M, Weisshaar B.2001. The R2R3-MYB gene family in Arabidopsis thaliana[J]. Current Opinion in Plant Biology, 4(5): 447-456.
19 Schlaeppi K, Aboumansour E,Buchala A, et al.2010. Disease resistance of Arabidopsis to Phytophthora brassicae is established by the sequential action of indole glucosinolates and camalexin[J]. Plant Journal, 62(5): 840-851.
20 Zhang N Q, Ho S C, Mo X F.2018. Glucosinolate and isothiocyanate intakes are inversely associated with breast cancer risk: A case-control study in China[J]. British Journal of Nutrition, 119(8): 957-964. |
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