|
|
|
| Cloning and Mutant Construction of Cold-upregulated Glycosyltransferase-like Gene (OsCUGT1) from Guizhou Landrace Rice 'Pingtang Heinuo' (Oryza sativa ssp. japonica) |
| CAI Ming-Liang1, CHEN Rong1, HUANG Xiao-Zhen1,2,*, ZHAO De-Gang1,3,* |
1 The Key Laboratory of Plant Resources Conservation and Germplasm Innovationin Mountainous Region (Ministry of Education), College of Life Sciences, Guizhou University, Guiyang 550025, China;
2 College of Tea Sciences, Guizhou University, Guiyang 550025, China;
3 Guizhou Academy of Agricultural Sciences, Guiyang Branch of DUS Center, Ministry of Agriculture and Rural Sciences, Guiyang 550006, China;
|
|
|
|
|
Abstract Plant glycosyltransferases (GTs) play an important role in regulating growth and stress adaptation. To further explore and study the biological functions of new glycosyltransferases in rice (Oryza sativa), in this study, a cold-upregulated glycosyltransferase-like gene 1 (OsCUGT1) was isolated and cloned, which was based on the cold treatment transcriptome library of Guizhou landrace rice 'Pingtang Heinuo' (O. sativa ssp. japonica). The results of qRT-PCR showed that the expression of OsCUGT1 gene was significantly induced by cold treatment. The subcellular localization results showed that it was localized in the chloroplasts. Phylogenetic analysis showed that OsCUGT1 was conserved in monocot plant species. Furthermore, sequence alignment revealed that there were several SNP sites between 'Pingtang Heinuo' and O. sativa ssp. Nipponbare in the OsCUGT1 gene. Among them, the site of 511 bp was changed from C to A, which consequently caused the 170 th amino acid to change from lysine (Q) to glutamine (K). Diversity analysis of the OsCUGT1 gene sequence revealed that there were 12 SNP sites of them were found in the exon region. To further explore the biological function of OsCUGT1, the OsCUGT1 gene was edited by CRISPR/Cas9 technique. The targeted editing mutants of oscugt1 were successfully obtained by optimizing Agrobacterium-mediated transformation method, screening of hygromycin selection and sequencing analysis. The results of this study can provide research materials and theoretical basis for further clarifying the role of OsCUGT1 in rice growth development and stress resistance．
|
|
Received: 06 September 2020
|
|
|
|
Corresponding Authors:
*xzhuang@gzu.edu.cn; dgzhao@gzu.edu.cn
|
|
|
|
[1] 陈璐. 2020. 拟南芥糖基转移酶基因UGT76F1和UGT71C3的功能及分子机制[D]. 博士学位论文, 山东大学, 导师: 侯丙凯, pp. 93-94.
(Chen L.2020. The function and molecular mechanism of Arabidopsis glycosyltransferase genes UGT76F1 and UGT71C3[D]. Thesis for Ph.D., Shandong University, Supervisor: Hou B K, pp. 93-94.)
[2] 丁杨林. 2015. 蛋白激酶OST1调控拟南芥响应低温胁迫的分子机制[D]. 博士学位论文, 中国农业大学, 导师: 杨淑华, pp. 36-48.
(Ding Y L.2015. The molecular mechanism of protein kinase OST1 regulating Arabidopsis response to low temperature stress[D]. Thesis for Ph.D., China Agricultural University, Supervisor: Yang S H, pp. 36-48.)
[3] 高璐, 薛永来. 2016. 农杆菌介导的水稻快速高效基因转化系统[J]. 江苏农业科学, 47(04): 39-42.
(Gao L, Xue Y L.2016. Agrobacterium-mediated rapid and efficient gene transformation system in rice[J]. Jiangsu Agricultural Sciences, 47(04): 39-42.)
[4] 黄小贞, 曾晓芳, 李建容, 等. 2017. 基于CRISPR/Cas9技术的水稻转录因子tify1a和tify1b突变体的创建与分析[J]. 农业生物技术学报, 25(06): 1003-1012.
(Huang X Z, Zeng X F. Li J R, et al.2017. Creation and analysis of mutants of rice transcription factors tify1a and tify1b based on CRISPR/Cas9 technology[J]. Journal of Agricultural Biotechnology, 25(06): 1003-1012.)
[5] 刘怒安, 曾晓芳, 赵德刚. 2019. 贵州禾来拢茎尖遗传转化体系的研究[J]. 种子, 38(06): 25-29.
(Liu N A, Zeng X F, Zhao D G.2019. Study on the genetic transformation system of Guizhou helairong stem apex[J]. Seed, 38(06): 25-29.)
[6] 孟祥州, 严菊, 邢宏堃, 等. 2015. 水稻NOL (NYC1-like)基因序列多样性、单倍型效应以及表达谱分析[J]. 分子植物育种, 13(03): 491-496.
(Meng X Z, Yan J, Xing H K, et al.2015. Sequence diversity, haplotype effect and expression profile analysis of rice NOL (NYC1-like) gene[J]. Molecular Plant Breeding, 13(03): 491-496.)
[7] 唐丽, 李曜魁, 张丹, 等. 2016. 基于基因组编辑技术的水稻靶向突变特征及遗传分析[J]. 遗传, 38(8): 746-755.
(Tang L, Li Y K, Zhang D, et al.2016. Characteristics and genetic analysis of targeted mutations in rice based on genome editing technology[J]. Inheritance, 38(8): 746-755.)
[8] 杨桥, 蔺自敏, 侯详文, 等. 2014. 小麦遗传转化中潮霉素筛选体系的建立及应用[J]. 麦类作物学报, 34(03): 304-310.
(Yang Q, Lin Z M, Hou X W, et al.2014. Establishment and application of hygromycin screening system in wheat genetic transformation[J]. Journal of Triticeae Crops, 34(03): 304-310.)
[9] 张笑寒, 仇志浪, 赵德刚. 2016. 农杆菌介导McCHIT1基因遗传转化水稻茎尖研究[J]. 中国农学通报, 32(27): 114-120.
(Zhang X H, Qiu Z L. Zhao D G.2016. Agrobacterium-mediated genetic transformation of rice stem tip with McCHIT1 gene[J]. Chinese Agricultural Science Bulletin, 32(27): 114-120.)
[10] Amos R A, Mohnen D.2019. Critical review of plant cell wall matrix polysaccharide glycosyltransferase activities verified by heterologous protein expression[J]. Frontiers in Plant Science, 915(10): 389-396.
[11] Anders N, Wilkinson M D, Lovegrove A, et al.2012. Glycosyl transferases in family 61 mediate arabinofuranosyl transfer onto xylan in grasses[J]. Proceedings of the National Academy of Sciences of the USA, 109(3): 989-993.
[12] Cao P J, Bartley L E, Jung K H, et al.2008. Construction of a rice glycosyltransferase phylogenomic database and identification of rice-diverged glycosyltransferases[J]. Molecular Plant, 1(5): 858-877.
[13] Chopra R, Simpson C E, Hillhouse A, et al.2018. SNP genotyping reveals major QTLs for plant architectural traits between A-genome peanut wild species[J]. Molecular Genetics and Genomics, 293(6): 1477-1491.
[14] Cui X, Wang Y, Wu J, et al.2019. The RNA editing factor DUA1 is crucial to chloroplast development at low temperature in rice[J]. The New Phytologist, 221(2): 834-849.
[15] Ellis M D, Hoak J M, Ellis B W, et al.2020. Quantitative Real-time pcr analysis of individual flue-cured tobacco seeds and seedlings reveals seed transmission of Tobacco mosaic virus[J]. Phytopathology, 110(1): 194-205.
[16] Fayos I, Mieulet D, Petit J, et al.2019. Engineering meiotic recombination pathways in rice[J]. Plant Biotechnology Journal, 17(11): 2062-2077.
[17] Guo H, Wu T, Li S, et al.2019. The methylation patterns and transcriptional responses to chilling stress at the seedling stage in rice[J]. International Journal of Molecular Sciences, 20(20): 5080-5089.
[18] Jang Y J, Seo M, Hersh C P, et al.2009. An evolutionarily conserved non-synonymous SNP in a leucine-rich repeat domain determines anthracnose resistance in watermelon[J]. Theoretical and Applied Genetics, 132(2): 473-488.
[19] Kilasi N L, Singh J, Vallejos C E, et al.2018. Heat stress tolerance in rice (Oryza sativa L.): Identification of quantitative trait loci and candidate genes for seedling growth under heat stress[J]. Frontiers in Plant Science, 9(15): 78-89.
[20] Kim W, Kim J Y, Cho S L, et al.2008. Glycosyltransferase: A specific marker for the discrimination of Bacillus anthracis from the Bacillus cereus group[J]. Journal of Medical Microbiology, 57(3): 279-286.
[21] Ko J H, Kim B G, Hur H G, et al.2006. Molecular cloning, expression and characterization of a glycosyltransferase from rice[J]. Plant Cell Reports, 25(7): 741-746.
[22] Lairson L L, Henrissat B, Davies G J, et al.2008. Glycosyltransferases: Structures, functions, and mechanisms[J]. Annual Review of Biochemistry, 77: 521-555.
[23] Lee H, Xiong L, Gong Z, et al.2001. The Arabidopsis HOS1 gene negatively regulates cold signal transduction and encodes a RING finger protein that displays cold-regulated nucleo-cytoplasmic partitioning[J]. Genes & Development, 15(7): 912-24.
[24] Lee K, Eggenberger A L, Banakar R, et al.2019. CRISPR/Cas9-mediated targeted T-DNA integration in rice[J]. Plant Molecular Biology, 99(4-5): 317-328.
[25] Li P, Li Y J, Wang B, et al.2017. The Arabidopsis UGT87A2, a stress-inducible family 1 glycosyltransferase, is involved in the plant adaptation to abiotic stresses[J]. Physiologia Plantarum, 159(4): 416-432.
[26] Lin J S, Huang X X, Li Q, et al.2016. UDP-glycosyltransferase 72B1 catalyzes the glucose conjugation of monolignols and is essential for the normal cell wall lignification in Arabidopsis thaliana[J]. The Plant Journal: For cell and Molecular Biology, 88(1): 26-42.
[27] Liu Q, Chen T T, Xiao D W, et al.2017. OsIAGT1 is a glucosyltransferase gene involved in the glucose conjugation of auxins in rice[J]. Rice (New York, N.Y.), 12(1): 92-123.
[28] Ma Y, Dai X, Xu Y, et al.2015. COLD1 confers chilling tolerance in rice[J]. Cell. 160(6): 1209-1221.
[29] Manishankar P, Kudla J.2015. Cold tolerance encoded in one SNP[J]. Cell. 160(6): 1045-1046.
[30] Rennie E A, Hansen S F, Baidoo E E, et al.2012. Three members of the Arabidopsis glycosyltransferase family 8 are xylan glucuronosyltransferases[J]. Plant Physiology, 159(4): 1408-1417.
[31] Song T, Zhang Q, Wang H, et al.2018. OsJMJ703, a rice histone demethylase gene, plays key roles in plant development and responds to drought stress[J]. Plant Physiology and Biochemistry, 132(10): 183-188.
[32] Shi Y, Gong Z.2015. One SNP in COLD1 determines cold tolerance during rice domestication[J]. Journal of Genetics and Genomics, 42(4): 133-134.
[33] Shi Y, Liu Y J, Cao S Y, et al.2020. Glycosyltransferase OsUGT90A1 helps protect the plasma membrane during chilling stress in rice[J]. Journal of Experimental Botany, 71(9): 2723-2739.
[34] Shi Y, Tian S, Hou L, et al.2012. Ethylene signaling negatively regulates freezing tolerance by repressing expression of CBF and type-A ARR genes in Arabidopsis[J]. The Plant Cell, 24(6): 2578-95.
[35] Vogt T, Jones P.2000. Glycosyltransferases in plant natural product synthesis: Characterization of a supergene family[J]. Trends in Plant Science, 5(9): 380-386.
[36] Wang M, Zhang T, Peng H, et al.2018. Rice premature leaf senescence 2, encoding a glycosyltransferase (gt), is involved in leaf senescence[J]. Frontiers in Plant Science, 26(9): 560-574.
[37] Wilson A E, Tian L.2019. Phylogenomic analysis of UDP-dependent glycosyltransferases provides insights into the evolutionary landscape of glycosylation in plant metabolism[J]. The Plant Journal: For Cell and Molecular Biology, 100(6): 1273-1288.
[38] Wu Z L, Person A D, Anderson M, et al.2018. Imaging specific cellular glycan structures using glycosyltransferases via click chemistry[J]. Glycobiology, 28(2): 69-79.
[39] Zhang B, Zhao T, Yu W, et al.2014. Functional conservation of the glycosyltransferase gene GT47A in the monocot rice[J]. Journal of Plant Research, 127(3): 423-432.
[40] Zhang Y, Li J, Gao C.2016. Generation of stable transgenic rice (Oryza sativa L.) by Agrobacterium-mediated transformation[J]. Current Protocols in Plant Biology, (2): 235-246.
[41] Zhou G K, Zhong R Q, Richardson Elizabeth A, et al.2006. The poplar glycosyltransferase GT47C is functionally conserved with Arabidopsis fragile fiber8[J]. Plant & Cell Physiology, 47(9): 1229-1240.
[42] Zhang H, Zhang J, Wei P, et al.2014. The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation[J]. Plant Biotechnology Journal, 12(6): 797-807.
[43] Zheng Y, Liao C, Zhao S, Wang C, et al.2017. The glycosyltransferase QUA1 regulates chloroplast-associated calcium signaling during salt and drought stress in Arabidopsis[J]. Plant & Cell Physiology, 58(2): 329-341.
[44] Zhou Y, Li S, Qian Q, et al.2009. BC10, a DUF266-containing and golgi-located type Ⅱ membrane protein, is required for cell-wall biosynthesis in rice (Oryza sativa L.)[J]. The Plant Journal: For Cell and Molecular Biology, 57(3): 446-462. |
|
|
|
