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作物学报 ›› 2023, Vol. 49 ›› Issue (4): 917-925.doi: 10.3724/SP.J.1006.2023.24064

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝型油菜SUMO蛋白家族成员鉴定及Bna.SUMO1.C08基因的功能研究

陈慧(), 肖清, 汪华栋, 文静, 马朝芝, 涂金星, 沈金雄, 傅廷栋, 易斌*()   

  1. 华中农业大学/作物遗传改良全国重点实验室/国家油菜工程技术研究中心, 湖北武汉 430070
  • 收稿日期:2022-03-22 接受日期:2022-07-21 出版日期:2023-04-12 网络出版日期:2022-08-12
  • 通讯作者: *易斌, E-mail: yibin@mail.hzau.edu.cn
  • 作者简介:E-mail: 15661644378@163.com
  • 基金资助:
    国家重点研发计划项目(2016YFD01008)

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 Published:2023-04-12 Published online:2022-08-12
  • Contact: *E-mail: yibin@mail.hzau.edu.cn
  • Supported by:
    National Key Research and Development Program of China(2016YFD01008)

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摘要：

蛋白翻译后修饰对蛋白的功能非常重要。SUMO化修饰就是一种非常重要的蛋白翻译后修饰, 它对植物生长发育的关键过程有很大的影响。甘蓝型油菜作为重要的油料和经济作物在SUMO化修饰方面却鲜有报道。为弥补这一空白, 本研究对甘蓝型油菜中的SUMO化修饰进行了探究。首先通过生物信息学方法在甘蓝型油菜中鉴定到31个SUMO蛋白成员, 分为3类: “典型”群组、“非典型”群组和SUMO-V。然后对甘蓝型油菜中AtSUMO1基因的同源基因Bna.SUMO1.C08进行表达模式分析, 发现该基因在根、叶和角果中表达比较高。亚细胞定位结果发现, Bna.SUMO1.C08蛋白定位于细胞核和内质网中。最后在甘蓝型油菜中过表达Bna.SUMO1.C08基因发现其能够增强植株对PEG胁迫的抵抗能力。本研究为后续甘蓝型油菜中SUMO化修饰的研究奠定了一定的基础。

关键词: 甘蓝型油菜, SUMO化修饰, Bna.SUMO1.C08

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

附表1

甘蓝型油菜SUMO蛋白家族成员基本信息表"

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

图1

甘蓝型油菜SUMO蛋白成员蛋白序列分析 A: 系统进化树分析。红色标注的Group 1为“典型”群组; 蓝色标注的Group 2为“非典型”群组; 紫色标注的Group 3为SUMO相关蛋白。B: Motif、Domain和基因结构分析。第1列为Motif; 第2列为Domain; 第3列为基因结构。C: 蛋白序列比对。红色长方体标注为β-grasp domain范围; 圆点标注为各作用位点。"

图2

甘蓝型油菜SUMO蛋白基因染色体定位和共线性分析 A: SUMO蛋白基因染色体位置可视化。B: 拟南芥、白菜、甘蓝和甘蓝型油菜物种物种内和物种间共线性分析。"

图3

Bna.SUMO1.C08表达模式分析和亚细胞定位 A: 表达模式分析。Bna.SUMO1.C08主要在根、叶和角果中表达。B: 亚细胞定位。Bna.SUMO1.C08定位于细胞核和内质网中。标尺: 1 μm。"

图4

过表达Bna.SUMO1.C08的转基因植株在mRNA水平上的鉴定 A: 过表达Bna.SUMO1.C08基因的载体结构图。B: 过表达植株Bna.SUMO1.C08的表达量检测。"

图5

PEG处理下过表达Bna.SUMO1.C08植株和野生型植株表型 A, B: 根长表型及统计。C, D: DAB染色和NBT染色。E, F: POD酶和SOD酶活性检测。*P < 0.05; **P < 0.01; ***P < 0.001。"

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