Identification and functional analysis of Euphorbia lathyris L. transcription factor ElWRI1
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摘要:
续随子（Euphorbia lathyris L.）种子可积累大量油脂且富含油酸，WRINKLED1（WRI1）为AP2/EREBP类转录因子，参与调控脂肪酸的合成及种子发育等生命过程。本研究通过对续随子的全基因组鉴定，获得了11个续随子ElWRI1转录因子基因，命名为ElWRI1-1~ElWRI1-11。结果显示，它们均具有两个AP2结构域；其编码蛋白均属于不稳定亲水蛋白，无跨膜结构；亚细胞定位预测显示所有WRI1蛋白均位于细胞核。表达分析结果表明，ElWRI1-6与ElWRI1-8在种子发育时期的表达量较高，且后者的表达模式与种子油脂积累模式一致。在本氏烟草（Nicotiana benthamiana L.）叶片中异源过表达ElWRI1-8，叶片中总油脂含量增加，且油酸含量显著上升。研究结果表明ElWRI1-8可能是调控续随子种子油脂和油酸生物合成的一个重要转录因子。
Abstract:Seeds of Euphorbia lathyris L. accumulate a substantial amount of oil enriched with oleic acid. The plant transcription factor WRINKLED1 (WRI1) is a member of the AP2/EREBP transcription factor family, which is involved in the regulation of fatty acid synthesis and seed development. Based on the whole genome data of E. lathyris, 11 ElWRI1 genes were identified, namely ElWRI1-1–ElWRI1-11, respectively. All identified ElWRI1s contained two AP2 domains. The ElWRI1 proteins were unstable hydrophilic proteins, lacking a transmembrane structure. The 11 ElWRI1 proteins were predicted to localize in the nucleus. Heatmap analysis revealed that ElWRI1-6 and ElWRI1-8 exhibited higher expression levels, with the expression pattern of ElWRI1-8 mirroring that of seed oil accumulation. Heterologous overexpression of ElWRI1-8 in Nicotiana benthamiana L. leaf tissues led to a significant increase in total oil and oleic acid contents in the leaf. This study indicates that ElWRI1-8 is an important transcription factor in regulating biosynthesis of oil and oleic acid in E. lathyris seeds, thus providing a new scientific basis for the high-yield and high-quality breeding of E. lathyris and the development of high-value oil products.
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Keywords:
- Euphorbia lathyris /
- Transcription factors /
- WRI1 /
- Oil synthesis /
- Leaf transient expression
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图 1 续随子 ElWRI1 基因的染色体定位
Figure 1. Chromosomal localization of WRI1 genes in Euphorbia lathyris
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图 5 WRI1 蛋白质序列对比
红色区域标记“VYL”3个氨基酸；黄色区域标记两个 AP2 结构域。
Figure 5. Sequence alignment of WRI1 proteins
“VYL” sequence present in first AP2 domain is marked by red box. Two AP2 domains are marked by black boxes.
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图 6 不同物种WRI1蛋白的系统发育分析
RcWRI1：蓖麻 (NP_001310691.1)；AtWRI1：拟南芥 (OAP01705.1)；JcWRI1：麻风树 (AIR74897.1)；HbWRI1：巴西橡胶树 (XP_021674385.1)；ZmWRI1：玉米 (ONM19989.1)；BnWRI1：甘蓝型油菜 (ADO16346.1)；CsWRI1：大豆 (KHN10130.1)；VvWRI1：葡萄 (XP_010659009.1)； CaWARI：辣椒(KAF3665339.1)； CiWRI1：山核桃 (QQP23243.1)； CnWRI1： 椰子 (KAG1365549.1)；TuWRI1：乌拉图小麦 (EMS52893.1)； GhWRI1：陆地棉(NP_001314053.1)；VmWRI1：皱果桐（APQ47387.1）。
Figure 6. Phylogenetic analysis of ElWRI1 among different plant species
RcWRI1: Ricinus communis (NP_001310691.1); AtWRI1: Arabidopsis thaliana (OAP01705.1); JcWRI1: Jatropha curcas (AIR74897.1); HbWRI1: Hevea brasiliensis (XP_021674385.1); ZmWRI1: Zea mays (ONM19989.1); BnWRI1: Brassica napus (ADO16346.1); CsWRI1: Clycine soja (KHN10130.1); VvWRI1: Vitis vinifera (XP_010659009.1); CaWARI: Capsicum annuum (KAF3665339.1); CiWRI1: Carya illinoinensis (QQP23243.1); CnWRI1: Cocos nucifera (KAG1365549.1); TuWRI1: Triticum urartu (EMS52893.1); GhWRI1: Gossypium hirsutum (NP_001314053.1); VmWRI1: Vernicia montana (APQ47387.1).
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图 7 续随子ElWRI1基因在种子发育不同时期的表达谱热图
作图数据使用为求导后的FPKM值（lgFPKM）。
Figure 7. Heatmap of ElWRI1 expression at different stages of seed development
Data are FPKM values after differentiation.
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图 8 续随子种子不同发育时期的油脂积累
Figure 8. Oil accumulation during seed development in Euphorbia lathyris
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图 9 过表达ElWRI1-8基因的烟草叶片总油脂含量
不同小写字母表示处理间差异显著（P < 0.05）。下同。
Figure 9. Total oil content in tobacco leaves overexpressing ElWRI1-8
Different lowercase letters indicate significant difference (P < 0.05). Same as below.
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[1] 中国科学院中国植物志编辑委员会. 中国植物志: 第44卷: 第3分册[M]. 北京: 科学出版社, 1997: 26-71. [2] 龚德勇,张燕,王晓敏,刘清国,李搏. 能源油料植物续随子的生物学及开发利用研究[J]. 江西农业学报,2011,23(8):39−41. doi: 10.3969/j.issn.1001-8581.2011.08.014 Gong DY,Zhang Y,Wang XM,Liu QG,Li B. Studies on biology,exploitation and utilization of energy oil plant Euphorbia lathyris[J]. Acta Agriculturae Jiangxi,2011,23 (8):39−41. doi: 10.3969/j.issn.1001-8581.2011.08.014
[3] Calvin M. Petroleum plantations for fuel and materials[J]. BioScience,1979,29 (9):533−538. doi: 10.2307/1307721
[4] 张卫明,史劲松,顾龚平. 生物质能的利用和能源植物的开发[J]. 南京师大学报(自然科学版),2007,30(3):68−74. doi: 10.3969/j.issn.1001-4616.2007.03.015 Zhang WM,Shi JS,Gu GP. Utilization of biomass energy and exploitation of energy plant[J]. Journal of Nanjing Normal University (Natural Science Edition)
,2007,30 (3):68−74. doi: 10.3969/j.issn.1001-4616.2007.03.015 [5] 刘冲,洪立洲,王茂文,丁海荣,朱小梅,等. 能源植物续随子及其研究进展[J]. 安徽农学通报,2011,17(23):119−120,134. doi: 10.3969/j.issn.1007-7731.2011.23.059 Liu C,Hong LZ,Wang MW,Ding HR,Zhu XM,et al. Progress of study on energy plant Euphorbia lathyris L.[J]. Anhui Agricultural Science Bulletin,2011,17 (23):119−120,134. doi: 10.3969/j.issn.1007-7731.2011.23.059
[6] 金梦阳,马冲,危文亮,冯顺山. 新型能源植物续随子的核型分析[J]. 中国油料作物学报,2007,29(2):213−215. doi: 10.3321/j.issn:1007-9084.2007.02.022 Jin MY,Ma C,Wei WL,Feng SS. Karyotype studies of new energy plant Euphorbia lathyris L.[J]. Chinese Journal of Oil Crop Sciences,2007,29 (2):213−215. doi: 10.3321/j.issn:1007-9084.2007.02.022
[7] 吉夏洁,郝青婷,刘宝玲,薛金爱,李润植. 蓖麻RcWRI1的鉴定及表达分析[J]. 山西农业大学学报(自然科学版),2017,37(4):229−234. doi: 10.3969/j.issn.1671-8151.2017.04.001 Ji XJ,Hao QT,Liu BL,Xue JA,Li RZ. Identification and expression analysis of the RcWRI1 in castor bean (Ricinus communis L. )[J]. Journal of Shanxi Agricultural University (Natural Science Edition)
,2017,37 (4):229−234. doi: 10.3969/j.issn.1671-8151.2017.04.001 [8] An D,Kim H,Ju S,Go YS,Kim HU,et al. Expression of Camelina WRINKLED1 isoforms rescue the seed phenotype of the Arabidopsis wri1 mutant and increase the triacylglycerol content in tobacco leaves[J]. Front Plant Sci,2017,8:229904.
[9] 陈贝贝. 大豆二酰甘油酰基转移酶（DGAT）和转录因子WRINKLED1（WRI1）功能研究[D]. 武汉: 华中农业大学, 2019: 1-20. [10] Maeo K,Tokuda T,Ayame A,Mitsui N,Kawai T,et al. An AP2-type transcription factor,WRINKLED1,of Arabidopsis thaliana binds to the AW-box sequence conserved among proximal upstream regions of genes involved in fatty acid synthesis[J]. Plant J,2009,60 (3):476−487. doi: 10.1111/j.1365-313X.2009.03967.x
[11] Shen B,Allen WB,Zheng PZ,Li CJ,Glassman K,et al. Expression of ZmLEC1 and ZmWRI1 increases seed oil production in maize[J]. Plant Physiol,2010,153 (3):980−987. doi: 10.1104/pp.110.157537
[12] Liu J,Hua W,Zhan GM,Wei F,Wang XF,et al. Increasing seed mass and oil content in transgenic Arabidopsis by the overexpression of wri1-like gene from Brassica napus[J]. Plant Physiol Biochem,2010,48 (1):9−15. doi: 10.1016/j.plaphy.2009.09.007
[13] Lim ARQ,Kong Q,Singh SK,Guo L,Yuan L,Ma W. Sunflower WRINKLED1 plays a key role in transcriptional regulation of oil biosynthesis[J]. Int J Mol Sci,2022,23 (6):3054. doi: 10.3390/ijms23063054
[14] 王志坤,常健敏,李丹丹,李文滨. 大豆GmWRI1a基因克隆及生物信息学分析[J]. 东北农业大学学报,2013,44(7):11−16. doi: 10.3969/j.issn.1005-9369.2013.07.003 Wang ZK,Chang JM,Li DD,Li WB. Cloning and bioinformatics analysis of GmWRI1a in soybean[J]. Journal of Northeast Agricultural University,2013,44 (7):11−16. doi: 10.3969/j.issn.1005-9369.2013.07.003
[15] 霍琳,及晓宇,王玉成. 农杆菌介导的烟草瞬时表达试验条件优化[J]. 分子植物育种,2016,14(1):80−85. doi: 10.13271/j.mpb.014.000080 Huo L,Ji XY,Wang YC. Transient expression conditions of tobacco transformation mediated by agrobacterium[J]. Molecular Plant Breeding,2016,14 (1):80−85. doi: 10.13271/j.mpb.014.000080
[16] 任国鹏,葛丽萍,孙超超,刘宝玲,李润植. 续随子二酰甘油酰基转移酶2基因(ElDGAT2)克隆与功能分析[J]. 植物生理学报,2019,55(8):1156−1166. Ren GP,Ge LP,Sun CC,Liu BL,Li RZ. Cloning and functional analysis of acyl-CoA:Diacylglycerol acyltransferase 2 gene (ElDGAT2) in Euphorbia lathyris[J]. Plant Physiol J,2019,55 (8):1156−1166.
[17] 王计平,张玲慧,赵静,王彦尊,李润植. 紫苏种子脂肪酸代谢及关键酶基因调控油脂合成规律的研究[J]. 中国粮油学报,2016,31(3):91−95. doi: 10.3969/j.issn.1003-0174.2016.03.017 Wang JP,Zhang LH,Zhao J,Wang YZ,Li RZ. Regulation of controlling oil synthesis by fatty acid metabolism of perilla seed and key enzyme gene[J]. Journal of the Chinese Cereals and Oils Association,2016,31 (3):91−95. doi: 10.3969/j.issn.1003-0174.2016.03.017
[18] 李璐. 紫苏种子脂肪酸代谢及合成相关基因PfPDAT的功能分析[D]. 晋中: 山西农业大学, 2017 : 1-30. [19] To A,Joubès J,Barthole G,Lécureuil A,Scagnelli A,et al. WRINKLED transcription factors orchestrate tissue-specific regulation of fatty acid biosynthesis in Arabidopsis[J]. Plant Cell,2012,24 (12):5007−5023.
[20] Filichkin SA,Priest HD,Givan SA,Shen RK,Bryant DW,et al. Genome-wide mapping of alternative splicing in Arabidopsis thaliana[J]. Genome Res,2010,20 (1):45−58. doi: 10.1101/gr.093302.109
[21] Ma W,Kong Q,Arondel V,Kilaru A,Bates PD,et al. WRINKLED1,a ubiquitous regulator in oil accumulating tissues from Arabidopsis embryos to oil palm mesocarp[J]. PLoS One,2013,8 (7):e68887. doi: 10.1371/journal.pone.0068887
[22] Krizek BA. AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain[J]. Nucleic Acids Res,2003,31 (7):1859−1868. doi: 10.1093/nar/gkg292
[23] Riechmann JL,Meyerowitz EM. The AP2/EREBP family of plant transcription factors[J]. Biol Chem,1998,379 (6):633−646. doi: 10.1515/bchm.1998.379.6.633
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