防风<i>SAUR32/23</i>基因生物信息学分析及编码蛋白的亚细胞定位

doi:10.7668/hbnxb.20194438

摘要/Abstract

摘要：

SAUR基因的生物信息学分析旨在揭示其结构、功能和进化关系,同时通过亚细胞定位,了解其在细胞中的位置,从而推断其生物学功能。这样分析有助于深入理解该基因在生物体内的作用机制和潜在应用,为调控防风种子萌发,缩短种子休眠时间和防风绿色栽培种植及育种提供重要理论和实践意义。研究进行了生物信息学分析,从防风克隆到2个基因SdSAUR32和SdSAUR23,分析这2个基因在防风种子不同休眠阶段的表现和亚细胞定位,以探究其生物学特征和功能。结果表明,SdSAUR32蛋白分子质量约为14.48 ku(pI为6.45),SdSAUR23蛋白分子质量约为10.29 ku(pI为8.00),均为亲水性蛋白;分析保守结构域的SdSAURs基因编码蛋白发现,SdSAUR32 和 SdSAUR23 的保守结构域相同,都属于由生长素诱导的超家族成员;进化分析表明,SdSAUR32和SdSAUR23基因编码蛋白均与胡萝卜的亲缘关系最近。亚细胞定位结果表明,防风SdSAUR32和SdSAUR23均定位于细胞核;防风种子的不同时期都有SdSAUR32和SdSAUR23的表达,但芽期的表达量显著高于休眠期和解除眠期,因此,推测SdSAUR32和SdSAUR23 可能在萌发和生长过程中起着重要作用。

关键词: 防风, SAUR, 生物信息学, 亚细胞定位

Abstract:

The bioinformatic analysis of SAUR genes aims to unveil their structure,function,and evolutionary relationships.Simultaneously,through subcellular localization,the goal was to understand their position within the cell,thereby inferring their biological functions.Such analyses contribute to a deeper understanding of the gene's mechanistic role and potential applications within an organism.This held particular theoretical and practical significance for regulating the germination of windproof seeds,reducing seed dormancy duration,and facilitating green cultivation and breeding of wind-resistant plants.In order to explore the biological characteristics and functions of SAUR genes,SdSAUR32 and SdSAUR23 were cloned from Saposhnikovia divaricata,and bioinformatics analysis was conducted to analyze the expression and subcellular localization of these two genes in different dormancy stages of Saposhnikovia divaricata seeds.The findings indicated that the SdSAUR32 protein had a molecular weight of approximately 14.48 ku(pI 6.45),while the SdSAUR23 protein had a molecular weight of around 10.29 ku(pI 8.00),confirming that both proteins were hydrophilic.The conserved domain analysis of SdSAURs gene encoded protein showed that SdSAUR32 and SdSAUR23 had the same conserved domain,which belonged to auxin induced superfamily members.The phylogenetic analysis showed that both SdSAUR32 and SdSAUR23 encoded protein was the closest relative to Daucus carota. The subcellular localization results indicated that both SdSAUR32 and SdSAUR23 were found within the nucleus.The SdSAUR32 and SdSAUR23 genes were found to be expressed during different stages of Saposhnikovia divaricata seed development.It was observed that the expression levels of these two genes were significantly higher during the germinal stage compared to the release dormancy stage and dormancy stage.This suggested that SdSAUR32 and SdSAUR23 may have a crucial role in the germination,growth,and development of Saposhnikovia divaricata seeds.This study offers not only insights for investigating the role of the SAUR gene family but also establishes atheoretical framework for examining the function of other genes in the parsnip family at the molecular level.

Key words: Saposhnikovia divaricata, SAUR, Bioinformatics, Subcellular localization

曹正林, 陈福源, 黄玉兰, 李有真, 丁洪霞, 夏凯旋, 魏文琦. 防风SAUR32/23基因生物信息学分析及编码蛋白的亚细胞定位[J]. 华北农学报, 2024, 39(2): 71-78. doi: 10.7668/hbnxb.20194438.

CAO Zhenglin, CHEN Fuyuan, HUANG Yulan, LI Youzhen, DING Hongxia, XIA Kaixuan, WEI Wenqi. Bioinformatics Analysis and Subcellular Localization of SAUR32/23 Genes in Saposhnikovia divaricata[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(2): 71-78. doi: 10.7668/hbnxb.20194438.

http://www.hbnxb.net/CN/Y2024/V39/I2/71

图/表 12

图1 SdSAURs基因扩增结果

Fig.1 The amplification outcome of the SdSAURs gene

图2 SdSAURs基因核苷酸及氨基酸序列

Fig.2 The nucleotide and amino acid sequences of the SdSAURs gene

图3 SdSAUR32的二级结构分析蓝色.α-螺旋;红色.延伸链;紫色.无规则卷曲;绿色.β-转角。图4同。

Fig.3 Second-structure analysis of SdSAUR32 Blue.α-helix; Red.Extended strand; Purple.Random coil; Green.β-turn.The same as Fig.4.

图4 SdSAUR23的二级结构分析

Fig.4 Second-structure analysis of SdSAUR23

图5 SdSAURs的三级级结构分析

Fig.5 3D structure analysis of SdSAURs

图6 SdSAUR32的保守结构域分析

Fig.6 Conserved domain analysis of SdSAUR32

图7 SdSAUR23的保守结构域分析

Fig.7 Conserved domain analysis of SdSAUR23

图8 SdSAUR32同源蛋白进化树分析

Fig.8 Phylogenetic analysis of SdSAUR32 gene and orthologs

图9 SdSAUR23同源蛋白进化树分析

Fig.9 Phylogenetic analysis of SdSAUR23 protein and orthologs

图10 SdSAURs基因的表达模式不同小写字母代表差异显著(P<0.05)。

Fig.10 Expression patterns of SdSAURs gene The different lowercase letters indicate that the difference is significant at 0.05 level.

图11 菌液PCR和质粒双酶切 A.pC2300-35S-GFP空载质粒;B.菌液PCR检测;C.双酶切验证。

Fig.11 Bacterial solution PCR and double enzyme digestion of plasmid A.pC2300-35S-GFP plasmid;B.PCR detection of bacterial liquid;C.Double digestion verification.

图12 SdSAURs亚细胞定位分析

Fig.12 Subcellular localization analysis of SdSAURs

参考文献 22

[1]	中国药典[S]. 一部.中国医药科技出版社.北京, 2020.
	Chinese Pharmacopoeia[S]. Part One.China Medical Science and Technology Press.Beijing, 2020.
[2]	柏桂顺, 王成功, 李江勇, 黎丽英, 姚姜羽, 巩丽虹. 中药防风活性成分及生理作用研究进展[J]. 中国民间疗法, 2020, 28(12):116-117.doi:10.19621/j.cnki.11-3555/r.2020.1252.
	Bai G S, Wang C G, Li J Y, Li L Y, Yao J Y, Gong L H. Research progress on active components and physiological functions of traditional Chinese medicine Saposhnikovia divaricata[J]. China's Naturopathy, 2020, 28(12):116-117.
[3]	刘双利, 姜程曦, 赵岩, 许永华, 王壮, 张连学. 防风化学成分及其药理作用研究进展[J]. 中草药, 2017, 48(10):2146-2152.doi:10.7501/j.issn.0253-2670.2017.10.032.
	Liu S L, Jiang C X, Zhao Y, Xu Y H, Wang Z, Zhang L X. Advance in study on chemical constituents of Saposhnikovia divaricate and their pharmacological effects[J]. Chinese Traditional and Herbal Drugs, 2017, 48(10):2146-2152.
[4]	赵文琦, 季兰. 种子休眠与解除机理的研究进展[J]. 山西农业科学, 2017, 45(3):477-481.doi:10.3969/j.issn.1002-2481.2017.03.38.
	Zhao W Q, Ji L. Research progress on mechanism of seed dormancy and dormancy breaking[J]. Journal of Shanxi Agricultural Sciences, 2017, 45(3):477-481.
[5]	Liu A H, Gao F, Kanno Y, Jordan M C, Kamiya Y, Seo M, Ayele B T. Regulation of wheat seed dormancy by after-ripening is mediated by specific transcriptional switches that induce changes in seed hormone metabolism and signaling[J]. PLoS One, 2013, 8(2):e56570.doi:10.1371/journal.pone.0056570. URL
[6]	宋松泉, 刘军, 唐翠芳, 张文虎, 徐恒恒, 张琪, 高家东. 生长素代谢与信号转导及其调控种子休眠与萌发的分子机制[J]. 科学通报, 2020, 65(34):3924-3943.doi:10.1360/TB-2020-0509.
	Song S Q, Liu J, Tang C F, Zhang W H, Xu H H, Zhang Q, Gao J D. Metabolism and signaling of auxins and their roles in regulating seed dormancy and germination[J]. Chinese Science Bulletin, 2020, 65(34):3924-3943.
[7]	Liu X D, Zhang H, Zhao Y, Feng Z Y, Li Q, Yang H Q, Luan S, Li J M, He Z H. Auxin controls seed dormancy through stimulation of abscisic acid signaling by inducing ARF-mediated ABI3 activation in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2013, 110(38):15485-15490.doi:10.1073/pnas.1304651110.
[8]	Wang S K, Bai Y H, Shen C J, Wu Y R, Zhang S N, Jiang D A, Guilfoyle T J, Chen M, Qi Y H. Auxin-related gene families in abiotic stress response in Sorghum bicolor[J]. Functional & Integrative, 2010, 10(4):533-546.doi:10.1007/s10142-010-0174-3.
[9]	Wu J, Liu S Y, He Y J, Guan X Y, Zhu X F, Cheng L, Wang J, Lu G. Genome-wide analysis of SAUR gene family in Solanaceae species[J]. Gene, 2012, 509(1):38-50.doi:10.1016/j.gene.2012.08.002. URL
[10]	Zhao J, Li W J, Sun S, Peng L L, Huang Z B, He Y Q, Wang Z F. The rice small auxin-up RNA gene OsSAUR33 regulates seed vigor via sugar pathway during early seed germination[J]. International Journal of Molecular Sciences, 2021, 22(4):1562.doi:10.3390/ijms22041562. URL
[11]	谢宏, 李舒文, 董迪, 王梦迪, 贾薇, 晁跃辉, 赵彦. 蒺藜苜蓿MtDWF1基因克隆、亚细胞定位及表达特征分析[J]. 草地学报, 2023, 31(4):984-991.doi:10.11733/j.issn.1007-0435.2023.04.006.
	Xie H, Li S W, Dong D, Wang M D, Jia W, Chao Y H, Zhao Y. Cloning,subcellular localization and expression pattern of MtDWF1 gene in Medicago truncatula[J]. Acta Agrestia Sinica, 2023, 31(4):984-991.
[12]	Guilfoyle T J, Hagen G. Auxin response factors[J]. Current Opinion in Plant Biology, 2007, 10(5):453-460.doi:10.1016/j.pbi.2007.08.014. pmid: 17900969
[13]	Jain M, Tyagi A K, Khurana J P. Genome-wide analysis,evolutionary expansion,and expression of early auxin-responsive SAUR gene family in rice(Oryza sativa)[J]. Genomics, 2006, 88(3):360-371.doi:10.1016/j.ygeno.2006.04.008. URL
[14]	Kant S, Bi Y M, Zhu T, Rothstein S J. SAUR39,a small auxin-up RNA gene,acts as a negative regulator of auxin synthesis and transport in rice[J]. Plant Physiology, 2009, 151(2):691-701.doi:10.1104/pp.109.143875. URL
[15]	左进城, 孙守梅, 王雅宁, 解卫海, 周瑞莲, 胡德昌. 赤霉素和吲哚乙酸对轮叶黑藻冬芽萌发和幼苗生长的作用[J]. 广东农业科学, 2014, 41(19):170-173.doi:10.16768/j.issn.1004-874x.2014.19.014.
	Zuo J C, Sun S M, Wang Y N, Xie W H, Zhou R L, Hu D C. Effects of GA and IAA on winter buds sprouting and seedling growth of Hydrilla verticillata[J]. Guangdong Agricultural Sciences, 2014, 41(19):170-173.
[16]	马正才, 马月芳. 不同浓度植物激素对青稞种子萌发及育苗的影响[J]. 甘肃高师学报, 2021, 26(5):14-19.doi:10.3969/j.issn.1008-9020.2021.05.005.
	Ma Z C, Ma Y F. Effects of the different concentration of plant hormones on the seed germination and seedling development of highland barley[J]. Journal of Gansu Normal Colleges, 2021, 26(5):14-19.
[17]	张素清, 陆爱君. 植物激素对种子萌发的影响[J]. 辽宁林业科技, 2015(3):59-61.doi:10.3969/j.issn.1001-1714.2015.03.022.
	Zhang S Q, Lu A J. Effects of plant hormones on seed germination[J]. Liaoning Forestry Science and Technology, 2015(3):59-61.
[18]	Chae K, Isaacs C G, Reeves P H, Maloney G S, Muday G K, Nagpal P, Reed J W. Arabidopsis SMALL AUXIN UP RNA63 promotes hypocotyl and stamen filament elongation[J]. The Plant Journal, 2012, 71(4):684-697.doi:10.1111/j.1365-313x.2012.05024.x.
[19]	Qiu T, Chen Y, Li M M, Kong Y Y, Zhu Y B, Han N, Bian H W, Zhu M Y, Wang J H. The tissue-specific and developmentally regulated expression patterns of the SAUR41 subfamily of small auxin up RNA genes:potential implications[J]. Plant Signaling & Behavior, 2013, 8(8):e25283.doi:10.4161/psb.25283.
[20]	Knauss S, Rohrmeier T, Lehle L. The auxin-induced maize gene ZmSAUR2 encodes a short-lived nuclear protein expressed in elongating tissues[J]. The Journal of Biological Chemistry, 2003, 278(26):23936-23943.doi:10.1074/jbc.m212585200. URL
[21]	Yang T B, Poovaiah B W. Molecular and biochemical evidence for the involvement of calcium/calmodulin in auxin action[J]. Journal of Biological Chemistry, 2000, 275(5):3137-3143.doi:10.1074/jbc.275.5.3137. pmid: 10652297
[22]	周大祥, 杨俊年, 龙泉洲, 殷幼平, 熊书. 青枯菌Ⅲ型分泌系统HrcN原核表达与亚细胞定位[J]. 西北农业学报, 2022, 31(9):1228-1234.doi:10.7606/j.issn.1004-1389.2022.09.016.
	Zhou D X, Yang J N, Long Q Z, Yin Y P, Xiong S. Prokaryotic expression and subcellular localization of HrcN protein in T3SS of Ralstonia solanacearum[J]. Acta Agriculturae Boreali-Occidentalis Sinica, 2022, 31(9):1228-1234.

选择文件类型/文献管理软件名称

选择包含的内容

防风SAUR32/23基因生物信息学分析及编码蛋白的亚细胞定位

Bioinformatics Analysis and Subcellular Localization of SAUR32/23 Genes in Saposhnikovia divaricata

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 12

参考文献 22

相关文章 15

编辑推荐

Metrics

本文评价

[1]	申金伟, 路建卫, 赵雪, 扎老, 成述儒, 梁春年. *牦牛DJ-1基因克隆、生物信息学及组织表达分析*[J]. 华北农学报, 2024, 39(2): 192-199.
[2]	姚妮, 罗仍卓么, 包斌武, 杨易, 王兴平. *肉牛ISG20基因编码区的克隆及其在子宫内膜上皮细胞容受态形成中的表达分析*[J]. 华北农学报, 2024, 39(2): 184-191.
[3]	胡莲, 刘益丽, 赵迪, 王泽宁, 江明锋. *麦洼牦牛MFSD4A基因克隆及组织表达分析*[J]. 华北农学报, 2024, 39(2): 174-183.
[4]	高泽仁, 潘鹏丞, 徐文文, 陈宝剑, 刘明君, 关志惠, 谢炳坤, 覃兆鲜. *陆川猪MyoD1基因克隆及组织表达分析*[J]. 华北农学报, 2024, 39(2): 168-173.
[5]	王慧珍, 张朝政, 黄一鸣, 黎耀心, 程子洋, 乐超银. *高粱抗病相关基因SbRPM1的克隆及表达分析*[J]. 华北农学报, 2024, 39(2): 153-160.
[6]	浦娜, 高巧丽, 王慧, 刘金朝, 陈嘉玲, 罗增铜, 肖武名, 陈淳. OsSAUR55通过GA途径调控水稻株型[J]. 华北农学报, 2024, 39(2): 106-115.
[7]	许华杰, 卢莉莉, 汤寓涵, 赵大球, 孟家松, 陶俊. *芍药PlSPL1基因克隆与表达特性分析*[J]. 华北农学报, 2024, 39(2): 55-61.
[8]	贺望兴, 李文金, 蒋咏梅, 童忠飞, 陈华玲, 李延升, 谢小群, 李琛. *灵芝LaeA基因的电子克隆、表达及生物信息学分析*[J]. 华北农学报, 2024, 39(2): 47-54.
[9]	张自阳, 周谦, 王依, 王志伟, 朱启迪, 茹振钢, 刘明久. *小麦倒春寒响应基因TaJAZ6的克隆、表达模式及亚细胞定位分析*[J]. 华北农学报, 2024, 39(1): 27-36.
[10]	袁惠君, 张瑞艳, 关玉晨, 余诗曼, 徐琰莹, 马倩国, 鲍婧婷. *扁果枸杞表皮蜡质转运相关基因LbABCG11的克隆及其表达特征分析*[J]. 华北农学报, 2023, 38(6): 45-54.
[11]	李明轩, 刘颖, 杨柏明, 张帆, 宋嘉欣, 粟柴静, 张卫华, 吴颖. 西瓜谷氨酸脱羧酶GADs的克隆与表达分析[J]. 华北农学报, 2023, 38(6): 18-25.
[12]	白璐, 辛翠花, 刘乐乐, 王俊杰, 简磊, 邵玉涛, 裴海霞, 郭江波. *本氏烟草NbEHD1基因的生物信息学分析、CRISPR/Cas9载体构建及遗传转化*[J]. 华北农学报, 2023, 38(5): 77-83.
[13]	黄渺, 罗悠悠, 倪芮, 肇瑾. *观赏辣椒ARP基因克隆及表达分析*[J]. 华北农学报, 2023, 38(5): 69-76.
[14]	张晓红, 许佩阳, 闫绪, 张贝贝, 于婵婵. *棉花GhFUL1基因及其启动子的克隆和功能分析*[J]. 华北农学报, 2023, 38(5): 60-68.
[15]	赖章龙, 阮超岭, 肖伟, 刘彦斌, 俞兆曦, 刘凯, 赛清云, 田永华, 张锋, 杨瑞兰, 柳婷, 杨立强, 王玉涛, 连总强. *兰州鲇NPY基因的克隆和表达分析*[J]. 华北农学报, 2023, 38(5): 227-238.

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

防风SAUR32/23基因生物信息学分析及编码蛋白的亚细胞定位

Bioinformatics Analysis and Subcellular Localization of SAUR32/23 Genes in Saposhnikovia divaricata

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 12

参考文献 22

相关文章 15

编辑推荐

Metrics

本文评价