Research article
Open Access
Overexpression of Brassica juncea wild‐type and mutant HMG‐CoA synthase 1 in Arabidopsis up‐regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance
Summary
Brassica juncea 3‐hydroxy‐3‐methylglutaryl‐CoA synthase (HMGS) is encoded by four isogenes (BjHMGS1‐BjHMGS4). In vitro enzyme assays had indicated that the recombinant BjHMGS1 H188N mutant lacked substrate inhibition by acetoacetyl‐CoA (AcAc‐CoA) and showed 8‐fold decreased enzyme activity. The S359A mutant demonstrated 10‐fold higher activity, while the H188N/S359A double mutant displayed a 10‐fold increased enzyme activity and lacked inhibition by AcAc‐CoA. Here, wild‐type and mutant BjHMGS1 were overexpressed in Arabidopsis to examine their effects in planta. The expression of selected genes in isoprenoid biosynthesis, isoprenoid content, seed germination and stress tolerance was analysed in HMGS overexpressors (OEs). Those mRNAs encoding enzymes 3‐hydroxy‐3‐methylglutaryl‐CoA reductase (HMGR), sterol methyltransferase 2 (SMT2), delta‐24 sterol reductase (DWF1), C‐22 sterol desaturase (CYP710A1) and brassinosteroid‐6‐oxidase 2 (BR6OX2) were up‐regulated in HMGS‐OEs. The total sterol content in leaves and seedlings of OE‐wtBjHMGS1, OE‐S359A and OE‐H188N/S359A was significantly higher than OE‐H188N. HMGS‐OE seeds germinated earlier than wild‐type and vector‐transformed controls. HMGS‐OEs further displayed reduced hydrogen peroxide (H2O2)–induced cell death and constitutive expression of salicylic acid (SA)‐dependent pathogenesis‐related genes (PR1, PR2 and PR5), resulting in an increased resistance to Botrytis cinerea, with OE‐S359A showing the highest and OE‐H188N the lowest tolerance. These results suggest that overexpression of HMGS up‐regulates HMGR, SMT2, DWF1, CYP710A1 and BR6OX2, leading to enhanced sterol content and stress tolerance in Arabidopsis.
Citing Literature
Number of times cited: 36
- Pan Liao, Xinjian Chen, Mingfu Wang, Thomas J. Bach and Mee‐Len Chye, Improved fruit α‐tocopherol, carotenoid, squalene and phytosterol contents through manipulation of Brassica juncea 3‐HYDROXY‐3‐METHYLGLUTARYL‐COA SYNTHASE1 in transgenic tomato, Plant Biotechnology Journal, 16, 3, (784-796), (2017).
- Tadao Wagatsuma, Eriko Maejima, Toshihiro Watanabe, Tomonobu Toyomasu, Masaharu Kuroda, Toshiya Muranaka, Kiyoshi Ohyama, Akifumi Ishikawa, Masami Usui, Shahadat Hossain Khan, Hayato Maruyama, Keitaro Tawaraya, Yuriko Kobayashi and Hiroyuki Koyama, Dark conditions enhance aluminum tolerance in several rice cultivars via multiple modulations of membrane sterols, Journal of Experimental Botany, 69, 3, (567), (2018).
- Miao Chen, Jingjing Chen, Na Luo, Rongda Qu, Zhenfei Guo and Shaoyun Lu, Cholesterol accumulation by suppression of SMT1 leads to dwarfism and improved drought tolerance in herbaceous plants, Plant, Cell & Environment, 41, 6, (1417-1426), (2018).
- Albert Ferrer, Teresa Altabella, Montserrat Arró and Albert Boronat, Emerging roles for conjugated sterols in plants, Progress in Lipid Research, 67, (27), (2017).
- Karla Ramirez-Estrada, Nídia Castillo, Juan A. Lara, Monserrat Arró, Albert Boronat, Albert Ferrer and Teresa Altabella, Tomato UDP-Glucose Sterol Glycosyltransferases: A Family of Developmental and Stress Regulated Genes that Encode Cytosolic and Membrane-Associated Forms of the Enzyme, Frontiers in Plant Science, 8, (2017).
- Junbo Du, Baolin Zhao, Xin Sun, Mengyuan Sun, Dongzhi Zhang, Shasha Zhang and Wenyu Yang, Identification and Characterization of Multiple Intermediate Alleles of the Key Genes Regulating Brassinosteroid Biosynthesis Pathways, Frontiers in Plant Science, 7, (2017).
- Qing Li, Gang Ding, Biao Li and Shun-Xing Guo, Transcriptome Analysis of Genes Involved in Dendrobine Biosynthesis in Dendrobium nobile Lindl. Infected with Mycorrhizal Fungus MF23 (Mycena sp.), Scientific Reports, 7, 1, (2017).
- , Characterization, Function, and Transcriptional Profiling Analysis of 3-Hydroxy-3-methylglutaryl-CoA Synthase Gene (GbHMGS1) towards Stresses and Exogenous Hormone Treatments in Ginkgo biloba, Molecules, 22, 10, (1706), (2017).
- Anup Kumar Singh, Sarma Rajeev Kumar, Varun Dwivedi, Avanish Rai, Shaifali Pal, Ajit K. Shasany and Dinesh A. Nagegowda, A WRKY transcription factor from Withania somnifera regulates triterpenoid withanolide accumulation and biotic stress tolerance through modulation of phytosterol and defense pathways, New Phytologist, 215, 3, (1115-1131), (2017).
- Rakesh Kumar, Abhishek Bohra, Arun K. Pandey, Manish K. Pandey and Anirudh Kumar, Metabolomics for Plant Improvement: Status and Prospects, Frontiers in Plant Science, 8, (2017).
- Pan Liao, Andréa Hemmerlin, Thomas J. Bach and Mee-Len Chye, The potential of the mevalonate pathway for enhanced isoprenoid production, Biotechnology Advances, 10.1016/j.biotechadv.2016.03.005, 34, 5, (697-713), (2016).
- Walter P. Suza and Joe Chappell, Spatial and temporal regulation of sterol biosynthesis in Nicotiana benthamiana, Physiologia Plantarum, 157, 2, (120-134), (2016).
- Lishia Teh and Christian Möllers, Genetic variation and inheritance of phytosterol and oil content in a doubled haploid population derived from the winter oilseed rape Sansibar × Oase cross, Theoretical and Applied Genetics, 10.1007/s00122-015-2621-y, 129, 1, (181-199), (2015).
- Zsofia Toth, Patrick Winterhagen, Balazs Kalapos, Yingcai Su, Laszlo Kovacs and Erzsebet Kiss, Expression of a Grapevine NAC Transcription Factor Gene Is Induced in Response to Powdery Mildew Colonization in Salicylic Acid-Independent Manner, Scientific Reports, 6, 1, (2016).
- Agata Lipko and Ewa Swiezewska, Isoprenoid generating systems in plants — A handy toolbox how to assess contribution of the mevalonate and methylerythritol phosphate pathways to the biosynthetic process, Progress in Lipid Research, 10.1016/j.plipres.2016.04.002, 63, (70-92), (2016).
- Jingjing Zhang, Lei Zhang, Jinkui Qiu and Hongjuan Nian, Isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis of Cryptococcus humicola response to aluminum stress, Journal of Bioscience and Bioengineering, 120, 4, (359), (2015).
- Anup Kumar Singh, Varun Dwivedi, Avanish Rai, Shaifali Pal, Sajjalavarahalli Gangireddy Eswara Reddy, Dodaghatta Krishnarao Venkata Rao, Ajit Kumar Shasany and Dinesh A. Nagegowda, Virus‐induced gene silencing of ithania somnifera squalene synthase negatively regulates sterol and defence‐related genes resulting in reduced withanolides and biotic stress tolerance, Plant Biotechnology Journal, 13, 9, (1287-1299), (2015).
- T. Blackmore, I. Thomas, R. McMahon, W. Powell and M Hegarty, Genetic–geographic correlation revealed across a broad European ecotypic sample of perennial ryegrass (Lolium perenne) using array-based SNP genotyping, Theoretical and Applied Genetics, 128, 10, (1917), (2015).
- N. Kusaira B. K. Ikram, Xin Zhan, Xi-Wu Pan, Brian C. King and Henrik T. Simonsen, Stable heterologous expression of biologically active terpenoids in green plant cells, Frontiers in Plant Science, 6, (2015).
- Majse Nafisi, Lorenzo Fimognari and Yumiko Sakuragi, Interplays between the cell wall and phytohormones in interaction between plants and necrotrophic pathogens, Phytochemistry, 112, (63), (2015).
- Shivani Saini, Isha Sharma and Pratap Kumar Pati, Versatile roles of brassinosteroid in plants in the context of its homoeostasis, signaling and crosstalks, Frontiers in Plant Science, 6, (2015).
- ZHI‐YAN DU, MO‐XIAN CHEN, QIN‐FANG CHEN, JI‐DONG GU and MEE‐LEN CHYE, Expression of rabidopsis acyl‐CoA‐binding proteins AtACBP1 and AtACBP4 confers Pb(II) accumulation in rassica juncea roots, Plant, Cell & Environment, 38, 1, (101-117), (2014).
- Pan Liao, Hui Wang, Andréa Hemmerlin, Dinesh A. Nagegowda, Thomas J. Bach, Mingfu Wang and Mee-Len Chye, Past achievements, current status and future perspectives of studies on 3-hydroxy-3-methylglutaryl-CoA synthase (HMGS) in the mevalonate (MVA) pathway, Plant Cell Reports, 33, 7, (1005), (2014).
- Arjun Sham, Ahmed Al-Azzawi, Salma Al-Ameri, Bassam Al-Mahmoud, Falah Awwad, Ahmed Al-Rawashdeh, Rabah Iratni, Synan AbuQamar and Raffaella Balestrini, Transcriptome Analysis Reveals Genes Commonly Induced by Botrytis cinerea Infection, Cold, Drought and Oxidative Stresses in Arabidopsis, PLoS ONE, 9, 11, (e113718), (2014).
- Pan Liao, Hui Wang, Mingfu Wang, An-Shan Hsiao, Thomas J. Bach, Mee-Len Chye and Joshua L. Heazlewood, Transgenic Tobacco Overexpressing Brassica juncea HMG-CoA Synthase 1 Shows Increased Plant Growth, Pod Size and Seed Yield, PLoS ONE, 9, 5, (e98264), (2014).
- Cécile Vriet, Eugenia Russinova and Christophe Reuzeau, From Squalene to Brassinolide: The Steroid Metabolic and Signaling Pathways across the Plant Kingdom, Molecular Plant, 6, 6, (1738), (2013).
- Wei Gao, Lu Long, Long-Fu Zhu, Li Xu, Wen-Hui Gao, Long-Qing Sun, Lin-Lin Liu and Xian-Long Zhang, Proteomic and Virus-induced Gene Silencing (VIGS) Analyses Reveal That Gossypol, Brassinosteroids, and Jasmonic acid Contribute to the Resistance of Cotton toVerticillium dahliae, Molecular & Cellular Proteomics, 12, 12, (3690), (2013).
- Ang Ren, Xiang Ouyang, Liang Shi, Ai-Liang Jiang, Da-Shuai Mu, Meng-Jiao Li, Qin Han and Ming-Wen Zhao, Molecular characterization and expression analysis of GlHMGS, a gene encoding hydroxymethylglutaryl-CoA synthase from Ganoderma lucidum (Ling-zhi) in ganoderic acid biosynthesis pathway, World Journal of Microbiology and Biotechnology, 29, 3, (523), (2013).
- Yuan Yuan, Jun Yu, Chao Jiang, Minhui Li, Shufang Lin, Xumin Wang and Luqi Huang, Functional Diversity of Genes for the Biosynthesis of Paeoniflorin and Its Derivatives in Paeonia, International Journal of Molecular Sciences, 14, 12, (18502), (2013).
- Andréa Hemmerlin, Post-translational events and modifications regulating plant enzymes involved in isoprenoid precursor biosynthesis, Plant Science, 203-204, (41), (2013).
- Clemens Krost, Romina Petersen, Stefanie Lokan, Bastienne Brauksiepe, Peter Braun and Erwin R. Schmidt, Evaluation of the hormonal state of columnar apple trees (Malus x domestica) based on high throughput gene expression studies, Plant Molecular Biology, 81, 3, (211), (2013).
- Moran Farhi, Magali Kozin, Shai Duchin and Alexander Vainstein, Metabolic engineering of plants for artemisinin synthesis, Biotechnology and Genetic Engineering Reviews, 29, 2, (135), (2013).
- Xiao Han, Cong Chen, Tae Kyung Hyun, Ritesh Kumar and Jae-Yean Kim, Metabolic module mining based on independent component analysis in Arabidopsis thaliana, Molecules and Cells, 34, 3, (295), (2012).
- Zuleika Briceño, Lorena Almagro, Ana B. Sabater-Jara, Antonio A. Calderón, María Angeles Pedreño and María Angeles Ferrer, Enhancement of phytosterols, taraxasterol and induction of extracellular pathogenesis-related proteins in cell cultures of Solanum lycopersicum cv Micro-Tom elicited with cyclodextrins and methyl jasmonate, Journal of Plant Physiology, 169, 11, (1050), (2012).
- Andréa Hemmerlin, John L. Harwood and Thomas J. Bach, A raison d’être for two distinct pathways in the early steps of plant isoprenoid biosynthesis?, Progress in Lipid Research, 10.1016/j.plipres.2011.12.001, 51, 2, (95-148), (2012).
- Tae Kyung Hyun, Yeonggil Rim, Hui-Jeong Jang, Cheol Hong Kim, Jongsun Park, Ritesh Kumar, Sunghoon Lee, Byung Chul Kim, Jong Bhak, Binh Nguyen-Quoc, Seon-Won Kim, Sang Yeol Lee and Jae-Yean Kim, De novo transcriptome sequencing of Momordica cochinchinensis to identify genes involved in the carotenoid biosynthesis, Plant Molecular Biology, 79, 4-5, (413), (2012).
