Welcome to Acta Agronomica Sinica，

Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (4): 917-925.doi: 10.3724/SP.J.1006.2023.24064

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Identification of SUMO protein family members and functional study of Bna.SUMO1.C08 gene in Brassica napus

CHEN Hui(), XIAO Qin, WANG Hua-Dong, WEN Jing, MA Chao-Zhi, TU Jin-Xing, SHEN Jin-Xiong, FU Ting-Dong, YI Bin*()   

  1. National Key Laboratory of Crop Genetic Improvement/National Engineering Research Center for Rapeseed/Huazhong Agricultural University, Wuhan 430070, Hubei, China
  • Received:2022-03-22 Accepted:2022-07-21 Online:2023-04-12 Published:2022-08-12
  • Contact: *E-mail: yibin@mail.hzau.edu.cn
  • Supported by:
    National Key Research and Development Program of China(2016YFD01008)

RichHTML409

35

PDF

321

Abstract:

Post-translational modifications are very important for protein function. SUMOylation is a very important post-translational modification, which has great influence on the key processes of plant growth and development. As an important oil and economic crops, there is few studies on SUMO modification in Brassica napus. To remedy this gap, SUMOylation in Brassica napus was investigated in this study. Firstly, 31 SUMO proteins were identified in Brassica napus by bioinformatics methods, which were divided into three groups (“canonical” SUMO, “non-canonical” SUMO, and SUMO-V). Secondly, the relative expression pattern indicated that, Bna.SUMO1.C08, a homologous gene of AtSUMO1, was highly expressed in roots, leaves, and silique. Subcellular localization revealed that Bna.SUMO1.C08 was localized in the nucleus and endoplasmic reticulum. Finally, Bna.SUMO1.C08 was overexpressed in Brassica napus and Bna.SUMO1.C08 could enhance plant resistance to PEG stress. This study laid a foundation for the subsequent research on SUMO modification in Brassica napus.

Key words: Brassica napus, SUMOylation, Bna.SUMO1.C08

Table S1

Basic information of SUMO protein family members in Brassica napus"

基因名
Gene name		 序列号
Sequence number		 基因长度
Gene length (bp)		 蛋白长度
Protein length (aa)		 分子量
Molecular weight (Da)		 等电点
Isoelectric point
Bna.SUMO1.C08 BnaC08T0170600ZS 304 101 11,083.39 4.91
Bna.SUMO1.A08 BnaA08T0169400ZS 304 101 11,124.43 4.91
Bna.SUMO1.C01 BnaC01T0209100ZS 316 105 11,589.98 4.93
Bna.SUMO1.A03 BnaA03T0494800ZS 304 101 11,123.50 4.91
Bna.SUMO1.A01 BnaA01T0162300ZS 303 100 11,120.40 4.93
Bna.SUMO1.C07 BnaC07T0472900ZS 295 98 10,889.23 4.91
Bna.SUMO1.A05 BnaA05T0335100ZS 259 86 10,309.86 10.61
Bna.SUMO1.C04 BnaC04T0237000ZS 256 85 9964.39 9.18
Bna.SUMO1.C06 BnaC06T0107300ZS 211 70 8117.07 9.25
Bna.SUMO1.C03 BnaC03T0634700ZS 211 70 8158.22 9.73
Bna.SUMO2.C09 BnaC09T0378000ZS 313 104 11,668.17 4.93
Bna.SUMO2.A10 BnaA10T0112600ZS 313 104 11,716.22 4.93
Bna.SUMO2.A02 BnaA02T0129200ZS 295 98 11,017.38 4.93
Bna.SUMO2.C02 BnaC02T0161400ZS 481 160 18,387.54 5.26
Bna.SUMO3.C03 BnaC03T0307000ZS 328 109 12,774.63 8.69
Bna.SUMO4.C07 BnaC07T0331100ZS 1213 404 46,871.13 9.15
Bna.SUMO4.A09 BnaA09T0050000ZS 244 81 9306.37 5.26
Bna.SUMO5.A04 BnaA04T0211000ZS 304 101 11,358.08 9.55
Bna.SUMO5.C03 BnaC03T0183400ZS 364 121 13,485.51 9.57
Bna.SUMO5.C04b BnaC04T0521300ZS 304 101 11,358.08 9.55
Bna.SUMO5.A05 BnaA05T0114400ZS 307 102 11,257.98 9.33
Bna.SUMO5.A03 BnaA03T0157500ZS 166 55 6198.18 9.52
Bna.SUMO5.A06 BnaA06T0124100ZS 325 108 12,451.37 8.66
Bna.SUMO5.C04c BnaC04T0143500ZS 295 98 10,885.65 9.61
Bna.SUMO5.C05 BnaC05T0152200ZS 313 104 11,998.78 8.69
Bna.SUMO5.C08 BnaC08T0244100ZS 292 97 11,140.70 5.04
Bna.SUMO5.C04a BnaC04T0143300ZS 307 102 11,279.03 9.73
Bna.SUMO6.A06 BnaA06T0362200ZS 1192 397 45,931.67 6.45
Bna.SUMO6.C09 BnaC09T0036100ZS 271 90 10,505.94 5.12
Bna.SUM-V.C06 BnaC06T0314000ZS 1042 347 38,854.96 5.23
Bna.SUM-V.A07 BnaA07T0276200ZS 724 241 27,254.87 5.32

Fig. 1

Sequence analysis of SUMO protein members in Brassica napus A: phylogenetic tree analysis. Group 1 in red is a “canonica” group. Group 2 in blue is a “non-canonica” group. Group 3 in purple is a sumo-related protein. B: the analyses of motif, domain, and gene structure. The first column is Motif; the second column is Domain; the third section is genetic structure. C: protein sequence alignment. The red cuboid is labeled as β-grasp domain range; dots are action sites."

Fig. 2

Chromosomal localization and collinearity analysis of SUMO protein gene in Brassica napus A: the chromosome location visualization of SUMO protein genes. B: the intra-species and interspecies collinearity analysis of Arabidopsis thaliana, Brassica oleracea, Brassica rapa, and Brassica napus."

Fig. 3

Relative expression pattern and subcellular localization of Bna.SUMO1.C08 genes A: the relative expression pattern. Bna.SUMO1.C08 is mainly expressed in roots, leaves, and scones. B: the subcellular localization. Bna.SUMO1.C08 locates in the nucleus and endoplasmic reticulum. Bar: 1 μm."

Fig. 4

Identification of Bna.SUMO1.C08 over expression transgenic lines at mRNA level A: the vector structure diagram of Bna.SUMO1.C08 overexpressed genes. B: the relative expression levels of Bna.SUMO1.C08 in overexpressed plants."

Fig. 5

Phenotypes of Bna.SUMO1.C08 over expression transgenic lines and wide type lines under PEG treatment A, B: the phenotype and statistics of root length. C, D: DAB staining and NBT staining. E, F: the enzyme activity detection of POD and SOD. *: P < 0.05; **: P < 0.01; ***: P < 0.001."

[1] Vierstra R D. The expanding universe of ubiquitin and ubiquitin-like uodifiers. Plant Physiol, 2012, 160: 2-14.
doi: 10.1104/pp.112.200667 pmid: 22693286
[2] Augustine R C, York S L, Rytz T C, Vierstra R D. Defining the SUMO system in maize: SUMOylation is up-regulated during endosperm development and rapidly induced by stress. Plant Physiol, 2016, 171: 2191-2210.
doi: 10.1104/pp.16.00353 pmid: 27208252
[3] Augustine R C, Vierstra R D. SUMOylation: re-wiring the plant nucleus during stress and development. Curr Opin Plant Biol, 2018, 45: 143-154.
doi: S1369-5266(17)30239-X pmid: 30014889
[4] Saracco S A, Miller M J, Kurepa J, Vierstra R D. Genetic analysis of SUMOylation in Arabidopsis: conjugation of SUMO1 and SUMO2 to nuclear proteins is essential. Plant Physiol, 2007, 145: 119-134.
pmid: 17644626
[5] Miura K, Lee J, Miura T, Hasegawa P M. SIZ1 controls cell growth and plant development in Arabidopsis through salicylic acid. Plant Cell Physiol, 2010, 51: 103-113.
doi: 10.1093/pcp/pcp171
[6] Miura K, Lee J, Gong Q, Ma S, Jin J B, Yoo C Y, Miura T, Sato A, Bohnert H J, Hasegawa P M. SIZ1 regulation of phosphate starvation-induced root architecture remodeling involves the control of auxin accumulation. Plant Physiol, 2011, 155: 1000-1012.
doi: 10.1104/pp.110.165191 pmid: 21156857
[7] Son G H, Park B S, Song J T, Seo H S. FLC-mediated flowering repression is positively regulated by sumoylation. J Exp Bot, 2014, 65: 339-351.
doi: 10.1093/jxb/ert383 pmid: 24218331
[8] Li X, Zhou S, Liu Z, Lu L, Dang H, Li H, Chu B, Chen P, Ma Z, Zhao S, Li Z, Nocker S, Ma F, Guan Q. Fine-tuning of SUMOylation modulates drought tolerance of apple. Plant Biotechnol J, 2022, 20: 903-919.
doi: 10.1111/pbi.13772 pmid: 34978131
[9] Wang F, Liu Y, Shi Y, Han D, Wu Y, Ye W, Yang H, Li G, Cui F, Wan S, Lai J, Yang C. SUMOylation stabilizes the transcription factor DREB2A to improve plant thermotolerance. Plant Physiol, 2020, 183: 41-50.
doi: 10.1104/pp.20.00080 pmid: 32205452
[10] Fang Q, Zhang J, Zhang Y, Fan N, van den Burg H A, Huang C. Regulation of aluminum resistance in Arabidopsis involves the SUMOylation of the zinc finger transcription factor STOP1. Plant Cell, 2020, 32: 3921-3938.
doi: 10.1105/tpc.20.00687
[11] Verma V, Srivastava A K, Gough C, Campanaro A, Srivastava M, Morrell R, Joyce J, Bailey M, Zhang C, Krysan P J, Sadanandom A. SUMO enables substrate selectivity by mitogen-activated protein kinases to regulate immunity in plants. Proc Natl Acad Sci USA, 2021, 118: e2021351118.
doi: 10.1073/pnas.2021351118
[12] Chen C, Chen H, Zhang Y, Thomas H R, Frank M H, He Y, Xia R. TBtools: an integrative toolkit developed for interactive analyses of big biological data. Mol Plant, 2020, 13: 1194-1202.
doi: S1674-2052(20)30187-8 pmid: 32585190
[13] Marchler-Bauer A, Bryant S H. CD-search: protein domain annotations on the fly. Nucleic Acids Res, 2004, 32: 327-331.
pmid: 15215404
[14] Gasteiger E. ExPASy: the proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Res, 2003, 31: 3784-3788.
doi: 10.1093/nar/gkg563 pmid: 12824418
[15] Horton P, Park K J, Obayashi T, Fujita N, Harada H, Adams- Collier C J, Nakai K. WoLF PSORT: protein localization predictor. Nucleic Acids Res, 2007, 35: W585-W587.
doi: 10.1093/nar/gkm259 pmid: 17517783
[16] Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol, 2016, 33: 1870-1874.
doi: 10.1093/molbev/msw054 pmid: 27004904
[17] Zhang H, Gao S, Lercher M J, Hu S, Chen W. EvolView, an online tool for visualizing, annotating and managing phylogenetic trees. Nucleic Acids Res, 2012, 40: W569-W572.
doi: 10.1093/nar/gks576
[18] Bailey T L, Boden M, Buske F A, Frith M, Grant C E, Clementi L, Ren J, Li W W, Noble W S. MEME Suite: tools for motif discovery and searching. Nucleic Acids Res, 2009, 37: W202-W208.
doi: 10.1093/nar/gkp335
[19] Yu J, Tehrim S, Zhang F, Tong C, Huang J, Cheng X, Dong C, Zhou Y, Qin R, Hua W, Liu S. Genome-wide comparative analysis of NBS-encoding genes between Brassica species and Arabidopsis thaliana. BMC Genomics, 2014, 15: 3.
doi: 10.1186/1471-2164-15-3
[20] Cheng F, Sun R, Hou X, Zheng H, Zhang F, Zhang Y, Liu B, Liang J, Zhuang M, Liu Y, Liu D, Wang X, Li P, Liu Y, Lin K, Bucher J, Zhang N, Wang Y, Wang H, Deng J, Liao Y, Wei K, Zhang X, Fu L, Hu Y, Liu J, Cai C, Zhang S, Zhang S, Li F, Zhang H, Zhang J, Guo N, Liu Z, Liu J, Sun C, Ma Y, Zhang H, Cui Y, Freeling M R, Borm T, Bonnema G, Wu J, Wang X. Subgenome parallel selection is associated with morphotype diversification and convergent crop domestication in Brassica rapa and Brassica oleracea. Nat Genet, 2016, 48: 1218-1224.
doi: 10.1038/ng.3634 pmid: 27526322
[21] Kurepa J, Walker J M, Smalle J, Gosink M M, Davis S J, Durham T L, Sung D, Vierstra R D. The small ubiquitin-like modifier (SUMO) protein modification system in Arabidopsis. J Biol Chem, 2003, 278: 6862-6872.
doi: 10.1074/jbc.M209694200
[22] Li Y, Wang G, Xu Z, Li J, Sun M, Guo J, Ji W. Organization and regulation of soybean SUMOylation system under abiotic stress conditions. Front Plant Sci, 2017, 8: 1458.
doi: 10.3389/fpls.2017.01458 pmid: 28878795
[23] Foyer C H, Noctor G. Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications. Antioxid Redox Signal, 2009, 11: 861-905.
doi: 10.1089/ars.2008.2177
[24] Sun X, Wang P, Jia X, Huo L, Che R, Ma F. Improvement of drought tolerance by overexpressing MdATG18a is mediated by modified antioxidant system and activated autophagy in transgenic apple. Plant Biotechnol J, 2018, 16: 545-557.
doi: 10.1111/pbi.12794
[25] Nadarajah K K. ROS homeostasis in abiotic stress tolerance in plants. Int J Mol Sci, 2020, 21: 5208.
doi: 10.3390/ijms21155208
[26] You J, Chan Z. ROS regulation during abiotic stress responses in crop plants. Front Plant Sci, 2015, 6: 1092.
doi: 10.3389/fpls.2015.01092 pmid: 26697045
[1] Wang Chu-Rui, Li Kai-Xiang, Zhao Zhi, Xiao Lu, Tang Guo-Yong, Zhao Zhi-Gang, Xu Liang, Du De-Zhi, Liu Hai-Dong. Development and application of KASP markers for functional sites of the early flowering gene BnCRY2 in Brassica napus L. spring rapeseed [J]. Acta Agronomica Sinica, 2026, 52(3): 708-721.
[2] Ma Yi-Na, Wu Xiao-Ming-Yu, Li Ou-Qi, Wang Yuan, Chen Li, Zhang Ying-Chuan, Zhao Lun, Wen Jing, Fu Ting-Dong, Shen Jin-Xiong. Functional analysis of the Bna-miR1040-EIF3A module in regulating flowering time in rapeseed (Brassica napus) [J]. Acta Agronomica Sinica, 2026, 52(2): 349-362.
[3] Lou Hong-Xiang, Xing Ren-Peng, Wang Bo, Wang Jing, Xu Zheng-Hua, Zhao Jie, Kuai Jie, Zhou Guang-Sheng. Effects of sowing date on the utilization efficiency of light and temperature resources and yield in Brassica napus in the middle reaches of the Yangtze River [J]. Acta Agronomica Sinica, 2026, 52(2): 539-551.
[4] WANG Bin, MENG Jiang-Yu, QIU Hao-Liang, HE Ya-Jun, QIAN Wei. Identification and expression pattern analysis of the BnaDUF579 gene family in Brassica napus [J]. Acta Agronomica Sinica, 2025, 51(8): 2100-2110.
[5] XIA Qi, GUO Ying, WANG Kun-Mei, WANG Si-Yi, JU Jian-Ye, PENG Ya-Wen, LIU Zhong-Song, XIA Shi-Tou. Correlation between salicylic acid and anthocyanins accumulation in seeds of different varieties in Brassica napus [J]. Acta Agronomica Sinica, 2025, 51(5): 1189-1197.
[6] ZHANG Qin, DAI Cheng, MA Chao-Zhi. Auxin response reporter gene transformation of Brassica napus and dynamic signal analysis of GUS in different tissues [J]. Acta Agronomica Sinica, 2025, 51(3): 667-675.
[7] SUN Cheng-Ming, ZHOU Xiao-Ying, CHEN Feng, ZHANG Wei, WANG Xiao-Dong, PENG Qi, GUO Yue, GAO Jian-Qin, HU Mao-Long, FU San-Xiong, ZHANG Jie-Fu. Functional analysis and prediction of long non-coding RNA (lncRNA) in the regulation of branch angle in Brassica napus L. [J]. Acta Agronomica Sinica, 2025, 51(3): 559-567.
[8] ZHUANG Yue-Ming, SONG Yi, CHEN Hang-Hang, LU Zhi-Feng, LIAO Shi-Peng, LI Xiao-Kun, CONG Ri-Huan, REN Tao, LU Jian-Wei. Effects of application of nitrogen on endophytic microbiota and seed quality in rapeseed (Brassica napus L.) seeds [J]. Acta Agronomica Sinica, 2025, 51(12): 3357-3368.
[9] HUANG Rong, ZHOU Qu-Chen, CHEN Chu-Ming, LUO Qian, YI Dong, DU Chang-Huan, HUANG Xiang-Yu, SHENG Feng, DU Xue-Zhu. Effects of BnNRT2.3-like overexpression on nitrogen uptake, utilization efficiency, and yield in Brassica napus L. [J]. Acta Agronomica Sinica, 2025, 51(12): 3184-3197.
[10] JU Jian-Ye, YANG Liu, CHEN Hao, KANG Lei, XIA Shi-Tou, LIU Zhong-Song. Single-nucleus transcriptome analysis reveals the cellular differentiation trajectories and molecular mechanisms underlying yellow seed coat formation in rapeseed [J]. Acta Agronomica Sinica, 2025, 51(11): 2860-2874.
[11] ZHANG Wen, LI Yu, WANG Chuang, SHI Lei, DING Guang-Da. Functional study of the phosphate transporter protein BnaPT48 in Brassica napus [J]. Acta Agronomica Sinica, 2025, 51(11): 2983-2995.
[12] ZHAO Hui-Xia, GUO Yan-Li, ZHENG Yu-Ling, HE Ling, CHEN Rui, WANG Shan-Shan, ZENG Chang-Li, ZOU Jun, SHEN Jin-Xiong, FU Ting-Dong, LIU Xiao-Yun, WAN He-Ping. Differential gene expression analysis of response to alkaline salt stress in Brassica napus L. [J]. Acta Agronomica Sinica, 2025, 51(11): 3105-3118.
[13] HU Zhi-Kang, SHU Yu, WANG Hui, YANG Ying-Ying, LIAO Jun-Yu, LIU Jia, CHENG Hong-Tao, GUO Chen, ZHANG Yuan-Yuan, LIU Sheng-Yi, HU Qiong, MEI De-Sheng, LI Chao. Comprehensive evaluation of alkaline tolerance in Brassica napus at the seedling stage [J]. Acta Agronomica Sinica, 2025, 51(10): 2681-2692.
[14] WANG Chen, HE Dan, YAO Min, QIU Ping, HE Xin, XIONG Xing-Hua, KANG Lei, LIU Zhong-Song, QIAN Lun-Wen. Transcriptome-based identification of flowering candidate genes and functional characterization of BnaCOR27 in Brassica napus [J]. Acta Agronomica Sinica, 2025, 51(10): 2693-2704.
[15] XU Lin-Shan, GAO Geng-Dong, WANG Yu, WANG Jia-Xing, YANG Ji-Zhao, WU Ya-Rui, ZHANG Xiao-Han, CHANG Ying, LI Zhen, XIE Xiong-Ze, GONG De-Ping, WANG Jing, GE Xian-Hong. Analysis of expression patterns of laccase gene family members in Brassica napus and their association with stem fracture resistance [J]. Acta Agronomica Sinica, 2025, 51(1): 134-148.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd