Please wait a minute...
浙江大学学报(理学版)  2023, Vol. 50 Issue (3): 367-377    DOI: 10.3785/j.issn.1008-9497.2023.03.015
生命科学     
龙眼UBP家族全基因组鉴定及其在外源胁迫下的表达分析
童宁,张春渝,许小琼,陈晓慧,申序,赖钟雄()
福建农林大学 园艺植物生物工程研究所,福建 福州 350002
Genome-wide identification and expression analysis of Dimocarpus longanUBP family and its expression analysis under drought stress
Ning TONG,Chunyu ZHANG,Xiaoqiong XU,Xiaohui CHEN,Xu SHEN,Zhongxiong LAI()
Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002,China
 全文: PDF(4667 KB)   HTML( 8 )
摘要:

泛素特异蛋白酶(ubiquitin-specific protease,UBP)是去泛素化酶六大家族中最大的,广泛存在于真核生物中,保守性很高。UBP可通过去泛素化作用调节各种生物的生长发育。为了解龙眼(Dimocarpus longan Lour.)DlUBP基因家族的生物学功能及其在龙眼体胚早期发生过程中的作用,基于第3代龙眼基因组数据库与转录组数据库,对DlUBP全基因组进行鉴定,分析其基因结构、蛋白质结构域、启动子顺式元件及构建系统进化树,并对DlUBP基因在脱落酸处理和干旱胁迫处理下的表达情况进行分析。结果显示,DlUBP家族共有18个成员,分为10个亚家族,其中最大的亚家族有4个成员;各成员的氨基酸数为369~1 117个,内含子数为2~31个;在DlUBP启动子序列中存在大量光响应元件、厌氧诱导响应元件、激素响应元件,随着处理时间的增加,在脱落酸处理下,多数DlUBP基因的表达量呈降低趋势,在干旱胁迫处理下,有不少DlUBP基因的表达量升高,推测DlUBP基因在龙眼体胚早期参与脱落酸的调控并对干旱胁迫有应激反应。

关键词: 龙眼UBP基因成员鉴定表达情况    
Abstract:

Ubiquitin-specific protease (UBP) is the largest among the six families of deubiquitinating enzymes. It is widely found in eukaryotes and is highly conserved. They regulate the growth and development of various organisms through deubiquitination. In order to understand the biological function of DlUBP gene family and its role during somatic embryogenesis of Dimocarpus longan Lour. In this study, we performed genome-wide identification of DlUBP gene family members; analysed their genes structure, protein structure, promoter cis-acting elements and constructed phylogenetic tree based on the third-generation longan genome database and transcriptome database; then analyzed the expression of DlUBP under ABA treatment and drought stress. The results showed that the DlUBP gene family contains 18 members, which are divided into 10 subfamilies, the largest subfamily has four members; the number of amino acids are between 369-1 117, and the number of introns are between 2-31; There are a large number of light response elements, anaerobic induction response elements, and hormone response elements in the DlUBP promoter sequence with a prolonged procecsing time, the expression level of many DlUBP members showed a decreasing trend under abscisic acid treatment, and the expression level of many DlUBP genes increased under drought stress. It is speculated that the DlUBP gene family participates in the regulation of abscisic acid during somatic embryogenesis of longan and responses to drought stress.

Key words: Dimocarpus longan    UBP gene    genome-wide identification    expression
收稿日期: 2021-11-16 出版日期: 2023-05-19
CLC:  S 667.2  
基金资助: 国家自然科学基金资助项目(31572088);福建省高原学科建设经费项目(102/71201801101);福建农林大学科技创新专项基金项目(CXZX2020027A)
通讯作者: 赖钟雄     E-mail: laizx01@163.com
作者简介: 童宁(1999—),ORCID:https://orcid.org/0000-0009-2826-5666,女,硕士研究生,主要从事果树分子生物技术研究.
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  
童宁
张春渝
许小琼
陈晓慧
申序
赖钟雄

引用本文:

童宁, 张春渝, 许小琼, 陈晓慧, 申序, 赖钟雄. 龙眼UBP家族全基因组鉴定及其在外源胁迫下的表达分析[J]. 浙江大学学报(理学版), 2023, 50(3): 367-377.

Ning TONG, Chunyu ZHANG, Xiaoqiong XU, Xiaohui CHEN, Xu SHEN, Zhongxiong LAI. Genome-wide identification and expression analysis of Dimocarpus longanUBP family and its expression analysis under drought stress. Journal of Zhejiang University (Science Edition), 2023, 50(3): 367-377.

链接本文:

https://www.zjujournals.com/sci/CN/10.3785/j.issn.1008-9497.2023.03.015        https://www.zjujournals.com/sci/CN/Y2023/V50/I3/367

基因名称正向引物反向引物
DlUBP2ACCATGAGAGGAGGCCACTACGCACGCACATAGGCATCGCTGAT
DlUBP8-1CCATCTGGCTCGGAATGACTCGAGAAGACGGCTGAGTGGTTCCA
DlUBP12-1ACGATGAGATGCTGGTGCCACAACTGCCTGTGCGTCCACTGTA
DlUBP12-2CCTGCTGGTGCTGTGGAGAATCTCCGCCATTTGTAACCGCCAAC
DlUBP17GGAGGAATGCGGTTGCCAGAACGTGGTGGTGGGAGAGAAACAG
DlUBP19ATCGCTTTCGGGCTGCTCCAGCACTTCTTGTCGCCAGGACTG
DlUBP22-2AGGTCACCATCGGATCGGTTGTCCTGAATGCTGCCACCAACTGT
DlUBP23GCAGCCATCAGCTCCATCTGTGTGGGACATGGGAACCGAGACTT
表1  实时荧光定量 PCR 引物
基因名称基因注释到拟南芥中的成员染色体氨基酸数/个分子质量/ku等电点
DlUBP2Dlo002772AtUBP2Chr11 004109.265.54
DlUBP4Dlo017141AtUBP4Chr736942.115.93
DlUBP5Dlo020549AtUBP5Chr9946105.495.86
DlUBP8-1Dlo025465AtUBP8Chr12895100.675.51
DlUBP8-2Dlo029798AtUBP8Chr1483295.095.24
DlUBP9Dlo032528AtUBP9Chr15930104.655.14
DlUBP12-1Dlo011562AtUBP12Chr51 117130.805.50
DlUBP12-2Dlo029835AtUBP12Chr141 072125.805.48
DlUBP12-3Dlo025482AtUBP12Chr12957112.465.48
DlUBP16Dlo013767AtUBP16Chr61 071116.606.11
DlUBP17Dlo016225AtUBP17Chr787596.868.88
DlUBP19Dlo000876AtUBP19Chr171079.136.00
DlUBP21Dlo027879AtUBP21Chr1367374.855.66
DlUBP22-1Dlo000463AtUBP22Chr179289.047.99
DlUBP22-2Dlo000824AtUBP22Chr155663.528.46
DlUBP23Dlo019901AtUBP23Chr9962105.189.32
DlUBP25Dlo009614AtUBP25Chr462068.628.86
DlUBP26Dlo010069AtUBP26Chr484794.937.84
表2  DlUBP全基因组蛋白质特性
图1  DlUBP基因在染色体上的定位
图2  DlUBP基因结构与保守motif分布图
图3  DlUBP家族成员的蛋白质保守结构域分析
图4  植物UBP家族的系统进化树分析
图5  DlUBP基因启动子顺式作用元件
图6  DlUBP基因在龙眼体胚发生各阶段的特异表达
图7  6个DlUBP基因家族成员在不同脱落酸处理时间下的表达情况不同小写字母表示在0.05水平上具有显著性差异。
图8  6个DlUBP基因家族成员在不同干旱胁迫处理时间下的表达情况不同小写字母表示在0.05水平上具有显著性差异。
1 陈雨晗, 张令强, 贺福初. 去泛素化酶与基因表达调控[J].生物化学与生物物理进展, 2014, 41(2): 126-138. DOI:10.3724/SP.J.1206.2013.00124
CHEN Y H, ZHANG L Q, HE F C. Deubiquitinating enzymes and gene expression regulation[J]. Progress in Biochemistry and Biophysics, 2014, 41(2): 126-138. DOI:10.3724/SP.J.1206.2013.00124
doi: 10.3724/SP.J.1206.2013.00124
2 刘文斌. 去泛素化酶研究进展[J]. 武汉轻工大学学报, 2016, 35(3): 1-13. DOI:10.3969/j.issn.2095-7386.2016.03.001
LIU W B. Research progress of deubiquitinating enzymes[J]. Journal of Wuhan University of Light Industry, 2016, 35(3): 1-13. DOI:10.3969/j.issn.2095-7386.2016.03.001
doi: 10.3969/j.issn.2095-7386.2016.03.001
3 REYES-TURCU F E, VENTII K H, WILKINSON K D. Regulation and cellular roles of ubiquitin-specificdeubiquitinating enzymes[J]. Annual Review of Biochemistry, 2009, 78: 363-397. DOI:10.1146/annurev.biochem.78.082307.091526
doi: 10.1146/annurev.biochem.78.082307.091526
4 REYES-TURCU F E, WILKINSON K D. Polyubiquitin binding and disassembly by deubiquitinating enzymes[J]. Chemical Reviews, 2009, 109(4): 1495-1508. DOI:10.1021/cr800470j
doi: 10.1021/cr800470j
5 HU M, LI P, LI M, et al. Crystal structure of aUBP-family deubiquitinating enzyme in isolation and in complex with ubiquitin aldehyde[J]. Cell, 2002, 111(7): 1041-1054. DOI:10.1016/S0092-8674(02)01199-6
doi: 10.1016/S0092-8674(02)01199-6
6 刘艳芬. 拟南芥泛素特异蛋白酶家族的功能分析[D]. 北京: 中国协和医科大学, 2007.
LIU Y F. Functional Analysis of Arabidopsis Ubiquitin-Specific Protease Family[D]. Beijing: Peking Union Medical College, 2007,
7 LIU Y F, WANG F, ZHANG H Y, et al. Functional characterization of the Arabidopsis ubiquitin-specific protease gene family reveals specific role and redundancy of individual members in development[J]. The Plant Journal, 2008, 55(5): 844-856. DOI:10.1111/j.1365-313X.2008.03557.x
doi: 10.1111/j.1365-313X.2008.03557.x
8 YAN N, DOELLING J H, FALBEL T G, et al. The ubiquitin-specific protease family from Arabidopsis AtUBP1 and 2 are required for the resistance to the amino acid analog canavanine[J]. Plant Physiology, 2000, 124(4): 1828-1843. DOI:10.1104/pp.124.4.1828
doi: 10.1104/pp.124.4.1828
9 XIA C, FALONG L, YUE L, et al. Ubiquitin-specific proteases UBP12 and UBP13 act in circadian clock and photoperiodic flowering regulation in Arabidopsis [J]. Plant Physiology, 2013, 162(2): 897-906. DOI:10.1104/pp.112.213009
doi: 10.1104/pp.112.213009
10 SACCO J J, COULSON J M, CLAGUE M J, et al. Emerging roles of deubiquitinases in cancer-associated pathways[J]. IUBMB Life, 2010, 62(2): 140-157. DOI:10.1002/iub.300
doi: 10.1002/iub.300
11 SCHWERTMAN P, BEKKER-JENSEN S, MAILAND N. Regulation of DNA double-strand break repair by ubiquitin and ubiquitin-like modifiers[J]. Nature Reviews Molecular Cell Biology, 2016, 17(6): 379-394. DOI:10.1038/nrm.2016.58
doi: 10.1038/nrm.2016.58
12 王梦媛, 谭震, 刘蔚晴, 等. 泛素特异性蛋白酶USP13调控小鼠单核巨噬细胞破骨分化的研究[C]// 2020年中华口腔医学会口腔生物医学专业委员会第十次全国口腔生物医学学术年会暨第六次全国口腔杰青优青论坛. 上海: 中华口腔医学会, 2020: 166-167.
WANG M Y, TAN Z, LIU W Q, et al. Ubiquitin-specific protease USP13 regulates the osteoclast differentiation of mouse monocytes and macrophages[C]// 2020 the 10th National Oral Biomedical Academic Annual Conference of the Oral Biomedical Professional Committee of the Chinese Stomatological Association and the 6th National Oral Biomedical Forum Paper Collection. Shanghai: Chinese Stomatological Association, 2020: 166-167.
13 RUIHUA W, YANRONG S, QIAG Z, et al. Genome-wide identification and characterization of the UBP gene family in Moso bambooPhyllostachys edulis)[J]. International Journal of Molecular Sciences, 2019, 20(17): 4309. DOI:10.3390/ijms20174309
doi: 10.3390/ijms20174309
14 MOON Y K, HONG J P, CHO Y C, et al. Structure and expression of OsUBP6, an ubiquitin-specific protease 6 homolog in rice (Oryza sativa L)[J]. Molecules and Cells, 2009, 28(5): 463-472. DOI:10.1007/s10059-009-0138-4
doi: 10.1007/s10059-009-0138-4
15 LIN Y L, MIN J M, LAI R L, et al. Genome-wide sequencing of longan (Dimocarpus longan Lour) provides insights into molecular basis of its polyphenol-rich characteristics[J]. GigaScience, 2017, 6(5): 1-14. doi:10.1093/gigascience/gix023
doi: 10.1093/gigascience/gix023
16 VERCRUYSSEN L, TOGNETTI V B, GONZALEZ N, et al. Growth regulating factor stimulates Arabidopsis chloroplast division, photosynthesis, and leaf longevity[J]. Plant Physiology, 2015, 167(3): 817-832. DOI:10.1104/pp.114.256180
doi: 10.1104/pp.114.256180
17 赖钟雄. 龙眼生物技术研究[M]. 福州:福建科学技术出版社, 2003: 18-22.
LAI Z X. Longan Biotechnology Research[M]. Fuzhou: Fujian Science and Technology Press, 2003: 18-22.
18 HUANG S X, GAO Y F, LIU J K, et al. Genome-wide analysis of WRKY transcription factors in Solanum lycopersicum [J]. Molecular Genetics and Genomics, 2012, 287(6): 495-513. DOI:10.1007/s00438-012-0696-6
doi: 10.1007/s00438-012-0696-6
19 李衍常. 泛素链和泛素化底物修饰位点特异性的定量蛋白质组学研究[D]. 北京: 中国人民解放军军事医学科学院, 2016.
LI Y C. Quantitative Proteomics Study on the Specificity of Ubiquitin Chain and Ubiquitinated Substrate Modification Sites[D]. Beijing: Chinese Academy of Military Medical Sciences, 2016.
20 YI P, XIA W, WU R C, et al. SRC-3 coactivator regulates cell resistance to cytotoxic stress via TRAF4-mediated p53 destabilization[J].Genes & Development, 2013, 27(3): 274-287. DOI:10.1101/gad.203760.112
doi: 10.1101/gad.203760.112
21 OH K H, YANG S W, PARK M, et al. Control of AIF-mediated cell death by antagonistic functions of CHIP ubiquitin E3 ligase and USP2 deubiquitinating enzyme[J]. Cell Death and Differentiation, 2011, 18(8): 1326-1336. DOI:10.1038/cdd.2011.3
doi: 10.1038/cdd.2011.3
22 孙倩倩, 李方周, 赵文会. 去泛素化酶与肿瘤[J]. 中国生物化学与分子生物学报, 2021, 37(2): 145-152. DOI:10.13865/j.cnki.cjbmb.2020.12.1537
SUN Q Q, LI F Z, ZHAO W H. Deubiquitinating enzymes and tumors[J]. Chinese Journal of Biochemistry and Molecular Biology, 2021, 37(2): 145-152. DOI:10.13865/j.cnki.cjbmb.2020.12.1537
doi: 10.13865/j.cnki.cjbmb.2020.12.1537
23 刘石娟, 秦宗燕, 王雪, 等. 植物去泛素化酶研究进展[J]. 植物生理学报, 2016, 52(8): 1135-1141. DOI:10.13592/j.cnki.ppj.2016.0213
LIU S J, QIN Z Y, WANG X, et al. Research progress of plant deubiquitinating enzymes[J]. Chinese Journal of Plant Physiology, 2016, 52(8): 1135-1141. DOI:10.13592/j.cnki.ppj.2016.0213
doi: 10.13592/j.cnki.ppj.2016.0213
24 孙莹, 林艺灵, 赵鹏程, 等. 龙眼类受体蛋白激酶CRK家族全基因组鉴定及表达调控分析[J]. 热带作物学报, 2019, 40(10): 1924-1937. DOI:10.3969/j.issn.1000-2561.2019.10.005
SUN Y, LIN Y L ZHAO P C, et al. Genome identification and expression regulation analysis of longan-like receptor protein kinase CRK family[J]. Chinese Journal of Tropical Crops, 2019, 40(10): 1924-1937. DOI:10.3969/j.issn.1000-2561.2019. 10.005
doi: 10.3969/j.issn.1000-2561.2019. 10.005
25 DOELLING J H, YANN, KUREPA J, et al. The ubiquitin‐specific protease UBP14 is essential for early embryo development in Arabidopsis thaliana [J]. The Plant Journal, 2001, 27(5): 393-405. DOI:10. 1046/j.1365-313X.2001.01106.x
doi: 10. 1046/j.1365-313X.2001.01106.x
26 AN Z C, LIU Y L, OU Y, et al. Regulation of the stability of RGF1 receptor by the ubiquitin-specific proteases UBP12/UBP13 is critical for root meristem maintenance[J]. PNAS, 2018, 115(5): 1123-1128. DOI:10.1073/pnas.1714177115
doi: 10.1073/pnas.1714177115
27 LEE H J, KIM M S, SHIN J M, et al. The expression patterns of deubiquitinating enzymes, USP22 and Usp22 [J]. Gene Expression Patterns, 2006, 6(3): 277-284. DOI:10.1016/j.modgep.2005. 07.007
doi: 10.1016/j.modgep.2005. 07.007
28 杨雪, 雒军, 王引权. 调控植物黄酮类化合物生物合成的MYB转录因子研究进展[J]. 甘肃中医药大学学报, 2018, 35(6): 77-81. DOI:10.16841/j.issn1003-8450.2018.06.17
YAN X, LUO J, WANG Y Q. Research progress of MYB transcription factor regulating the biosynthesis of plant flavonoids[J]. Journal of Gansu University of Traditional Chinese Medicine, 2018, 35(6): 77-81. DOI:10.16841/j.issn1003-8450.2018.06.17
doi: 10.16841/j.issn1003-8450.2018.06.17
29 ZHAO J F, ZHOU H P, ZHANG M, et al. Ubiquitin-specific protease24 negatively regulates abscisic acid signalling in Arabidopsis thaliana [J]. Plant, Cell & Environment, 2016, 39(2): 427-440. DOI:10.1111/pce.12628
doi: 10.1111/pce.12628
30 DU Y T, ZHAO M J, WANG C T, et al. Identification and characterization of GmMYB118 responses to drought and salt stress[J]. BMC Plant Biology, 2018, 18(1): 320. DOI:10.1186/S12870-018-1551-7
doi: 10.1186/S12870-018-1551-7
[1] 刘生财, 段俊朋, 姚德恒, 林玉玲, 赖钟雄. 响应面试验优化超声波辅助提取龙眼愈伤组织中单宁的工艺[J]. 浙江大学学报(理学版), 2019, 46(1): 58-64.