马铃薯StNF-YA9基因的生物信息学及其表达分析

Bioinformatics and Expression Analysis of the StNF-YA9 Gene in Potato

摘要

NF-Y是一种普遍存在于真核生物中的转录因子，通常以NF-YA/B/C异源三聚体的形式发挥作用，其中NF-YA转录因子通过特异性结合DNA启动子区域的CCAAT box，调控下游靶基因表达以应对胁迫。通过前期对马铃薯NF-Ys基因家族的分析，发现沉默马铃薯StNF-YA9基因，马铃薯对干旱胁迫更为敏感。因此，从马铃薯品种‘Atlantic’中克隆了StNF-YA9基因，生物信息学分析表明该基因总长度为1 231 bp，CDS区长723 bp。通过构建StNF-YA9亚细胞定位融合表达载体确定其定位在细胞核中；采用qRT-PCR分析该基因组织和逆境胁迫下表达模式，结果表明其在叶片中相对表达量最高，芽中相对表达量最低，受到聚乙二醇（Polyethylene glycol，PEG）和脱落酸（Abscisic acid，ABA）诱导上调表达。研究结果为马铃薯StNF-YA9基因后续功能研究提供理论基础。

Abstract

NF-Y is a transcription factor ubiquitous in eukaryotes, typically functioning as NF-YA/B/C heterotrimers. The NF-YA transcription factor regulates the expression of downstream target genes in response to stress by specifically binding to the CCAAT box in the DNA promoter region. It was discovered that the potato exhibits increased sensitivity to drought stress when the StNF-YA9 gene is silenced through a comprehensive analysis of the potato NF-Ys gene family. In this research, the StNF-YA9 gene was cloned from the potato variety 'Atlantic'. Bioinformatics analysis showed that the total length of StNF-YA9 gene was 1 231 bp, and the CDS region length was 723 bp. The subcellular localization fusion expression vector of StNF-YA9 was constructed to determine its localization in the nucleus. The expression pattern of StNF-YA9 in potato tissues and under stress was analyzed by qRT-PCR. The results showed that the relative expression of the StNF-YA9 gene was highest in leaves and lowest in buds. The expression was up-regulated by polyethylene glycol (PEG) and abscisic acid (ABA). The results would provide the theoretical basis for subsequent functional studies on the potato StNF-YA9 gene.

关键词

马铃薯 / StNF-YA9 / 克隆 / 生物信息学分析 / 亚细胞定位

Key words

potato / StNF-YA9 / cloning / bioinformatics analysis / subcellular localization

参考文献

[1] 罗其友, 刘洋, 高明杰, 等. 中国马铃薯产业现状与前景[J]. 农业展望, 2015, 11(3): 35-40.

[2] 王芳, 刘雁南, 赵文. 推进中国马铃薯主食化进程研究[J]. 世界农业, 2016(3): 11-14.

[3] 徐宁, 张洪亮, 张荣华, 等. 中国马铃薯种植业现状与展望[J]. 中国马铃薯, 2021, 35(1): 81-96.

[4] Eiasu B K, Hammes P S, Sersderma P. Response of potato (Solanum tuberosum) tuber yield components to gel-polymer soil amendments and irrigation regimes[J]. New Zealand Journal of Crop and Horticultural Science, 2007, 817(3): 121-130.

[5] 周维博. 干旱半干旱地域提高灌区水资源综合效益研究进展与思考[J]. 干旱区资源与环境, 2003(5): 91-96.

[6] Mahajan S, Tuteja N. Cold, salinity and drought stresses: an overview[J]. Archives of Biochemistry and Biophysics, 2005, 444(2): 139-158.

[7] 陈兰兰, 王丽, 吴亚娟, 等. 植物响应干旱胁迫的分子和微生态机制[J/OL]. 分子植物育种, 1-15[2024-03-20]. http://kns.cnki.net/kcms/detail/46.1068.S.20230406.1634.006.html.

[8] Romier C, Cocchiarella F, Mantovani R, et al. The NF-YB/NF-YC structure gives insight into DNA binding and transcription regulation by CCAAT factor NF-Y[J]. Journal of Biological Chemistry, 2003, 278(2): 1336-1345.

[9] Thirumurugan T, Ito Y, Kubo T, et al. Identification, characterization and interaction of HAP family genes in rice[J]. Molecular Genetics and Genomics, 2008, 279(3): 279-289.

[10] Stephenson T J, Mcintre C L, Collet C, et al. Genome-wide identification and expression analysis of the NF-Y family of transcription factors in Triticum aestivum[J]. Plant Molecular Biology, 2007, 65(1-2): 77-92.

[11] Li S, Li K, Ju Z, et al. Genome-wide analysis of tomato NF-Y factors and their role in fruit ripening[J]. BMC Genomics, 2016, 17: 36.

[12] Li S, Zhang N, Zhu X, et al. Genome-wide analysis of NF-Y genes in potato and functional identification of StNF-YC9 in drought tolerance[J]. Frontiers in Plant Science, 2021, 12: 749688.

[13] Hackenberg D, Wu Y, Voigt A, et al. Studies on differential nuclear translocation mechanism and assembly of the three subunits of the Arabidopsis thaliana transcription factor NF-Y[J]. Molecular Plant, 2012, 5(4): 876-888.

[14] 徐珍珍, 季为, 陈祥龙, 等. 异常棉NF-YA家族基因鉴定及其在干旱胁迫下的表达分析[J]. 江苏农业学报, 2023, 39(6): 1265-1274.

[15] 王高升, 王佩瑶, 孙念礼, 等. BnaNF-YA2增强甘蓝型油菜渗透胁迫抗性的功能鉴定[J/OL]. 中国油料作物学报, 1-14[2024-03-20]. https://doi.org/10.19802/j.issn.1007-9084.2024038.

[16] 王霄, 魏涵, 李世贵, 等. 马铃薯休眠解除相关基因StNF-YA8的表达分析[J]. 农业生物技术学报, 2024, 32(1): 50-59.

[17] 王凯彤. 马铃薯StTCP15基因在块茎休眠解除过程中的功能研究[D]. 兰州: 甘肃农业大学, 2022.

[18] 王芳芳. 马铃薯StERF109基因的功能鉴定[D]. 兰州: 甘肃农业大学, 2021.

[19] Tang X, Zhang N, Si H, et al. Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress[J]. Plant Methods, 2017, 13(1): 85.

[20] Sukunari N V, Mishra A, Kumar A, et al. Revisiting the role of transcription factors in coordinating the defense response against citrus bark cracking viroid infection in commercial hop (Humulus lupulus L.)[J]. Viruses, 2019, 11(5): 421-433.

[21] Wang F, Chen H, Li Q, et al. GmWRKY27 interacts with GmMYB174 to reduce expression of GmNAC29 for stress tolerance in soybean plants[J]. Plant Journal, 2015, 83(2): 224-236.

[22] Albani D, Robert L S. Cloning and characterization of a Brassica napus gene encoding a homologue of the B subunit of a heteromeric CCAAT-binding factor[J]. Gene, 1995, 167(1-2): 209-213.

[23] Ma X, Zhu X, Li C, et al. Overexpression of wheat NF-YA10 gene regulates the salinity stress response in Arabidopsis thaliana[J]. Plant Physiology and Biochemistry, 2015, 86: 34-43.

[24] Wang B, Li Z, Ran Q, et al. ZmNF-YB16 overexpression improves drought resistance and yield by enhancing photosynthesis and the antioxidant capacity of maize plants[J]. Frontiers in Plant Science, 2018, 9: 709.

[25] Lee D K, Kim H I, Jang G, et al. The NF-YA transcription factor OsNF-YA7 confers drought stress tolerance of rice in an abscisic acid independent manner[J]. Plant Science, 2015, 241: 199-210.

[26] 宋波涛, 涂卫, 王海波, 等. 马铃薯种质演化与分类研究进展[J]. 中国马铃薯, 2023, 37(6): 536-554.

[27] 郑子凡, 于佳乐, 聂虎帅, 等. 干旱胁迫对18个马铃薯品种生理特性的影响[J]. 中国马铃薯, 2023, 37(4): 306-319.

[28] 王金丽, 杜晨曦, 周华坤, 等. 外源ABA与植物非生物胁迫抗逆机制[J]. 安徽农业科学, 2019, 47(13): 12-15.

[29] 思星如, 陈馨, 魏涵, 等. 马铃薯StWRKY57基因的生物信息学分析及亚细胞定位[J]. 中国马铃薯, 2023, 37(5): 385-396.

基金资助

国家重点研发计划项目(2022YFD1602103)

甘肃省科技重大专项计划项目(23ZDNA006)