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毛竹早期光诱导蛋白基因克隆及功能分析

娄永峰 高志民

娄永峰, 高志民. 毛竹早期光诱导蛋白基因克隆及功能分析[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200237
引用本文: 娄永峰, 高志民. 毛竹早期光诱导蛋白基因克隆及功能分析[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200237
LOU Yongfeng, GAO Zhimin. Cloning and functional analysis of early light induced protein genes of Phyllostachys edulis[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200237
Citation: LOU Yongfeng, GAO Zhimin. Cloning and functional analysis of early light induced protein genes of Phyllostachys edulis[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200237

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毛竹早期光诱导蛋白基因克隆及功能分析

doi: 10.11833/j.issn.2095-0756.20200237
基金项目: 国家自然科学基金资助项目(31971736,31370588)
详细信息
    作者简介: 娄永峰,从事林木遗传育种研究。E-mail: louyf1983@163.com
    通信作者: 高志民,研究员,从事竹藤生长发育的分子基础研究。E-mail: gaozhimin@icbr.ac.cn
  • 中图分类号: S722.3;Q781

Cloning and functional analysis of early light induced protein genes of Phyllostachys edulis

  • 摘要:   目的  探究早期光诱导蛋白(ELIP)基因在竹子光保护中的作用,为进一步阐述竹子光保护机制提供参考依据。  方法  以毛竹Phyllostachys edulis实生苗为材料,在前期研究的基础上克隆毛竹ELIP基因,利用qRT-PCR技术研究其在不同光照诱导下的表达谱,同时通过在拟南芥Arabidopsis thaliana中异位表达对1个基因的功能进行初步鉴定。  结果  克隆获得了3个毛竹ELIP基因(PeELIP1、PeELIP2和PeELIP3),分别编码165、179和182个氨基酸。蛋白结构分析表明:3个PeELIPs蛋白均具有典型的捕光叶绿素a/b结合蛋白功能域,含3个α-螺旋跨膜结构,属于叶绿素a/b结合蛋白超家族。序列比对及进化分析表明:PeELIPs与水稻Oryza sativa、玉米Zea mays等单子叶植物的ELIPs相似性较高,同源性达72%以上,聚类在同一分支。qRT-PCR分析表明:3个PeELIPs基因在毛竹黄化苗中仅检测到微弱表达,光照处理使3个基因的表达量均显著上调;同时在正常毛竹实生苗叶片中,随着光照强度的增强和强光胁迫处理时间的延长,3个PeELIPs基因的表达量都显著上调。过表达PeELIP3可减缓转基因拟南芥在强光下Fv/Fm的下降幅度,但未影响转基因植株的非光化学猝灭系数。  结论  毛竹中至少存在3个PeELIPs,且其表达均受光照的诱导。过量表达PeELIP3能够减缓转基因拟南芥受光抑制的程度,具有一定的光保护作用。图8表1参40
  • 图  1  毛竹与拟南芥、水稻、玉米的ELIPs氨基酸序列比对

    At:拟南芥Arabidopsis thaliana;Pe:毛竹Phyllostachys edulis;Os:水稻Oryza sativa;Zm:玉米Zea mays。TM1~TM3表示3个α-螺旋跨膜结构。叶绿素a/b结合蛋白超家族的保守域用红线标出,三角形符号表示叶绿素结合位点

    Figure  1  Alignment of the deduced amino acid sequences of ELIPs from Ph. edulis, A. thaliana, O. sativa and Z. mays

    图  2  基因ELIPs氨基酸序列构建的系统进化树

    At. 拟南芥Arabidopsis thaliana;Br. 芜青Brassica rapa;Cr. 莱茵衣藻Chlamydomonas reinhardtii;Ds. 杜氏盐藻Dunaliella salina;Gb. 银杏Ginkgo biloba;Gm. 大豆Glycine max;Hv. 大麦Hordeum vulgare;Mf. 黄花苜蓿Medicago falcate;Ms. 紫花苜蓿Medicago sativa;Os. 水稻Oryza sativa;Pe. 毛竹Phyllostachys edulis;Pp. 小立碗藓Physcomitrella patens;Ps. 豌豆Pisum sativum;Pt. 毛果杨Populus trichocarpa;Sc. 齿肋赤藓Syntrichia caninervis;Si. 番茄Solanum lycopersicum;Sr. 山齿藓Syntrichia ruralis;Ta. 普通小麦Triticum aestivum;Tp. 红车轴草Trifolium pretense;Zm. 玉米Zea mays

    Figure  2  Phylogenetic tree based on the amino acid sequences of ELIPs

    图  3  PeELIPs在毛竹黄化苗中的表达分析

    Figure  3  Expression analysis of PeELIPs in etiolated seedlings of moso bamboo

    图  4  不同光照强度处理下PeELIPs的表达分析

    Figure  4  Expression analysis of PeELIPs under different light intensity

    图  5  强光(1200 µmol·m−2·s−1)胁迫下PeELIPs的表达分析

    Figure  5  Expression profile analysis of PeELIPs under high light stress (1200 µmol·m−2·s−1)

    图  6  PeELIP3转基因拟南芥PCR鉴定(A)及表达检测(B)

    Figure  6  Detection of transgenic Arabidopsis by PCR (A) and expression analysis of PeELIP3 in transgenic lines (B)

    图  7  强光(1200 µmol·m−2·s−1)对PeELIP3转基因拟南芥Fv/Fm的影响

    Figure  7  Effect of high light (1200 µmol·m−2·s−1) on Fv/Fm of PeELIP3 transgenic Arabidopsis

    图  8  PeELIP3转基因拟南芥非光化学猝灭分析

    Figure  8  Analysis of NPQ in PeELIP3 transgenic Arabidopsis

    表  1  引物序列

    Table  1.   Sequence of primers

    用途基因名称正向引物序列(5′→3′)反向引物序列(5′→3′)
    基因克隆PeELIP1ATGGCGACCAAGGTGGCCTTCTAGACGTTGACGAGCGGGGC
    PeELIP2ATGGCGACGACCATGATGGCTTACACTACTAGTTTTAGACGTTGAC
    PeELIP3ATGGCGACGACCATGATGACTTAGATGTTGACGAACGGCGC
    表达分析PeELIP1ATCATGTCCGCTGACGCCGACTTTGTGCTAGACGTTGACGAGC
    PeELIP2ACGACCATGATGGCCTCGAGTTGGGCGTCTCCGTTGGATC
    PeELIP3GCGCATCTAGCCTGTGCAATTTGTTCTGGGCCCTCACGAC
    下载: 导出CSV
  • [1] HEDDAD M, ADAMSKA I. The evolution of light stress proteins in photosynthetic organisms [J]. Comp Funct Genom, 2002, 3(6): 504 − 510. doi:  10.1002/cfg.221
    [2] BECK J, LOHSCHEIDER J N, ALBERT S, et al. Small one-helix proteins are essential for photosynthesis in Arabidopsis [J]. Front Plant Sci, 2017, 1(8): 7. doi:  10.3389/fpls.2017.00007.
    [3] MEYER G, KLOPPSTECH K. A rapidly light-induced chloroplast protein with a high turnover coded for by pea unclear DNA [J]. Eur J Biochem, 1984, 138(1): 201 − 207. doi:  10.1111/j.1432-1033.1984.tb07900.x
    [4] GRIMM B, KRUSE E, KLOPPSTECH K. Transiently expressed early light-inducible thylakoid proteins share transmembrane domains with light-harvesting chlorophyll binding proteins [J]. Plant Mol Biol, 1989, 13(5): 583 − 593. doi:  10.1007/BF00027318
    [5] ENGELKEN J, BRINKMANN H, ADAMSKA I. Taxonomic distribution and origins of the extended LHC (light-harvesting complex) antenna protein superfamily [J]. BMC Evol Biol, 2010, 10: 233. doi:  10.1186/1471-2148-10-233
    [6] HEDDAD M, NORÉN H, REISER V, et al. Differential expression and localization of early light-induced proteins in Arabidopsis [J]. Plant Physiol, 2006, 142(1): 75 − 87. doi:  10.1104/pp.106.081489
    [7] PINTO F, BERTI M, OLIVARES D, et al. Leaf development, temperature and light stress control of the expression of early light-inducible proteins (ELIPs) in Vitis vinifera L. [J]. Environ Exp Bot, 2011, 72(2): 278 − 283. doi:  10.1016/j.envexpbot.2011.04.002
    [8] ZHUO Chunliu, CAI Jiongliang, GUO Zhenfei. Overexpression of early light-induced protein (ELIP) gene from Medicago sativa ssp. falcata increases tolerance to abiotic stresses [J]. Agron J, 2013, 105(5): 1433 − 1440. doi:  10.2134/agronj2013.0155
    [9] WANG Huanli, CAO Fuliang, LI Guangping, et al. The transcript profiles of a putative early light-induced protein (ELIP) encoding gene in Ginkgo biloba L. under various stress conditions [J]. Acta Physiol Plant, 2015, 37(1): 1720. doi:  10.1007/s11738-014-1720-8
    [10] TIMERBAEV V, DOLGOV S. Functional characterization of a strong promoter of the early light-inducible protein gene from tomato [J]. Planta, 2019, 250(4): 1307 − 1323. doi:  10.1007/s00425-019-03227-x
    [11] HUTIN C, NUSSAUME L, MOISE N, et al. Early light-induced proteins protect Arabidopsis from photooxidative stress [J]. PNAS, 2003, 100(8): 4921 − 4926. doi:  10.1073/pnas.0736939100
    [12] BERTI M, PINTO M. Expression of early light induced protein in grapevine and pea, under different conditions and its relation with photoinhibition [J]. Chil J Agr Res, 2012, 72(3): 371 − 378. doi:  10.4067/S0718-58392012000300011
    [13] ZENG Qin, CHEN Xinbo, WOOD A J. Two early light-inducible protein (ELIP) cDNAs from the resurrection plant Tortula ruralis are differentially expressed in response to desiccation, rehydration, salinity and high light [J]. J Exp Bot, 2002, 53(371): 1197 − 1205. doi:  10.1093/jexbot/53.371.1197
    [14] SÄVENSTRAND H, OLOFSSON M, SAMUELSSON M, et al. Induction of early light-inducible protein gene expression in Pisum sativum after exposure to low levels of UV-B irradiation and other environmental stresses [J]. Plant Cell Rep, 2004, 22(7): 532 − 536. doi:  10.1007/s00299-003-0743-1
    [15] SHIMOSAKA E, SASANUMA T, HANDA H. A wheat cold-regulated cDNA encoding an early light-inducible protein (ELIP): its structure, expression and chromosomal location [J]. Plant Cell Physiol, 1999, 40(3): 319 − 325. doi:  10.1093/oxfordjournals.pcp.a029544
    [16] 周雅, 张道远. 植物早期光诱导蛋白的结构、功能及表达模式研究进展[J]. 基因组学与应用生物学, 2015, 34(6): 1339 − 1346.

    ZHOU Ya, ZHANG Daoyuan. The research progress of early light induced protein in plant [J]. Genom Appl Biol, 2015, 34(6): 1339 − 1346.
    [17] RIZZA A, BOCCACCINI A, LOPEZ-VIDRIERO I, et al. Inactivation of the ELIP1 and ELIP2 genes affects Arabidopsis seed germination [J]. New Phytol, 2011, 190(4): 896 − 905. doi:  10.1111/j.1469-8137.2010.03637.x
    [18] 杜澜, 谢锦忠, 赖秋香, 等. 遮荫对绿竹容器苗光合作用及生长的影响[J]. 生态学杂志, 2019, 38(1): 67 − 73.

    DU Lan, XIE Jinzhong, LAI Qiuxiang, et al. The effects of shading on photosynthetic characteristics and growth of Dendrocalamopsis oldhami seedling in contaniner [J]. Chin J Ecol, 2019, 38(1): 67 − 73.
    [19] 周哲宇, 徐超, 胡策, 等. 毛竹快速生长期的叶绿素荧光参数特征[J]. 浙江农林大学学报, 2018, 35(1): 75 − 80. doi:  10.11833/j.issn.2095-0756.2018.01.010

    ZHOU Zheyu, XU Chao, HU Ce, et al. Chlorophyll fluorescence characteristics of Phyllostachys edulis during its fast growth period [J]. J Zhejiang A&F Univ, 2018, 35(1): 75 − 80. doi:  10.11833/j.issn.2095-0756.2018.01.010
    [20] JIANG Zhehui, PENG Zhenhua, GAO Zhimin, et al. Characterization of different isoforms of the light-harvesting chlorophyll a/b complexes of photosystem Ⅱ in bamboo [J]. Photosynth Res, 2012, 50(1): 129 − 138. doi:  10.1007/s11099-012-0009-7
    [21] 娄永峰. 毛竹光保护及相关基因功能研究[D]. 北京: 中国林业科学研究院, 2016.

    LOU Yongfeng. Study on Photoprotection and Related Gene Function of Phyllostachys edulis [D]. Beijing: Chinese Academy of Forestry, 2016.
    [22] ZHAO Hansheng, LOU Yongfeng, SUN Huayu, et al. Transcriptome and comparative gene expression analysis of Phyllostachys edulis in response to high light [J]. BMC Plant Biol, 2016, 16: 34. doi:  10.1186/s12870-016-0720-9
    [23] GAO Zhimin, LIU Qing, ZHENG Bo, et al. Molecular cloning and functional analysis of violaxanthin deepoxidase gene (PeVDE) from Phyllostachys edulis [J]. Plant Cell Rep, 2013, 32(9): 1381 − 1391. doi:  10.1007/s00299-013-1450-1
    [24] LOU Yongfeng, SUN Huayu, LI Lichao, et al. Characterization and primary functional analysis of a bamboo ZEP gene from Phyllostachys edulis [J]. DNA Cell Biol, 2017, 36(9): 747 − 758. doi:  10.1089/dna.2017.3705
    [25] LOU Yongfeng, SUN Huayu, WANG Sining, et al. Expression and functional analysis of two PsbS genes in bamboo (Phyllostachys edulis) [J]. Physiol Plantarum, 2018, 163(4): 459 − 471. doi:  10.1111/ppl.12690
    [26] PENG Zhenhua, LU Ying, LI Lubin, et al. The draft genome of the fast-growing non-timber forest species moso bamboo (Phyllostachys heterocycla) [J]. Nat Genet, 2013, 45(4): 456 − 461. doi:  10.1038/ng.2569
    [27] KUMAR S, STECHER G, TAMURA K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets [J]. Mol Biol Evol, 2016, 33(7): 1870 − 1874. doi:  10.1093/molbev/msw054
    [28] LIVAK K J, SCHMITTGEN D T. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method [J]. Methods, 2001, 25(4): 402 − 408. doi:  10.1006/meth.2001.1262
    [29] FAN Chunjie, Ma Jinmin, Guo Qirong, et al. Selection of reference genes for quantitative real-time PCR in bamboo (Phyllostachys edulis)[J]. PLoS One, 2013, 8(2): e56573. doi:  10.1371/journal.pone.0056573.
    [30] CLOUGH S J, BENT A F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana [J]. Plant J, 1998, 16(6): 735 − 743. doi:  10.1046/j.1365-313x.1998.00343.x
    [31] 韩志国. 20种湿地植物的叶绿素荧光特性[D]. 广州: 暨南大学, 2006.

    HAN Zhiguo. Chlorophyll Fluorescence of 20 Species of Wetland Plants[D]. Guangzhou: Jinan University, 2006.
    [32] LI Xianwen, LIU Huijuan, XIE Suxia, et al. Isolation and characterization of two genes of the early light-induced proteins of Camellia sinensis [J]. Photosynthetica, 2013, 51: 305 − 311. doi:  10.1007/s11099-013-0025-2
    [33] AHRAZEM O, ARGANDOÑA J, CASTILLO R, et al. Identification and cloning of differentially expressed SOUL and ELIP genes in saffron stigmas using a subtractive hybridization approach[J]. PLoS One, 2016, 11(12): e0168736. doi:  10.1371/journal.pone.0168736.
    [34] PENG Yanhui, LIN Wuling, WEI Hui, et al. Phylogenetic analysis and seasonal cold acclimation-associated expression of early light-induced protein genes of Rhododendron catawbiense [J]. Physiol Plantarum, 2008, 132(1): 44 − 52.
    [35] 李菲, 张习敏, 王野影, 等. 复苏植物旋蒴苣苔早期光诱导蛋白的特征分析[J]. 基因组学与应用生物学, 2019, 38(3): 1162 − 1167.

    LI Fei, ZHANG Ximin, WANG Yeying, et al. Characterization analysis of early light-induced proteins (ELIPs) in resuscitation plant B. hygrometrica [J]. Genom Appl Biol, 2019, 38(3): 1162 − 1167.
    [36] BRUNO A K, WETZEL C M. The early light-inducible protein (ELIP) gene is expressed during the chloroplast-to-chromoplast transition in ripening tomato fruit [J]. J Exp Bot, 2004, 55(408): 2541 − 2548. doi:  10.1093/jxb/erh273
    [37] 陈晨. 盐生杜氏藻Dscbr基因的光保护功能及机制研究[D]. 成都: 四川大学, 2007.

    CHEN Chen. Studies on Photoprotective Function and Mechanism of Dscbr Gene in the Green Alga Dunaliella salina[D]. Chengdu: Sichuan University, 2007.
    [38] GOSS R, LEPETIT B. Biodiversity of NPQ [J]. J Plant Physiol, 2015, 172: 13 − 32. doi:  10.1016/j.jplph.2014.03.004
    [39] 李先文, 谢素霞, 张苏锋, 等. 植物早期光诱导蛋白基因研究进展[J]. 植物生理学报, 2011, 47(6): 540 − 544.

    LI Xianwen, XIE Suxia, ZHANG Sufeng, et al. Research advances in the genes of early light-induced proteins of plants [J]. J Plant Physiol, 2011, 47(6): 540 − 544.
    [40] TZVETKOVA-CHEVOLLEAU T, FRANCK F, ALAWADY A E, et al. The light stress-induced protein ELIP2 is a regulator of chlorophyll synthesis in Arabidopsis thaliana [J]. Plant J, 2007, 50(5): 795 − 809. doi:  10.1111/j.1365-313X.2007.03090.x
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出版历程
  • 收稿日期:  2020-03-26
  • 修回日期:  2020-04-09

毛竹早期光诱导蛋白基因克隆及功能分析

doi: 10.11833/j.issn.2095-0756.20200237
    基金项目:  国家自然科学基金资助项目(31971736,31370588)
    作者简介:

    娄永峰,从事林木遗传育种研究。E-mail: louyf1983@163.com

    通信作者: 高志民,研究员,从事竹藤生长发育的分子基础研究。E-mail: gaozhimin@icbr.ac.cn
  • 中图分类号: S722.3;Q781

摘要:   目的  探究早期光诱导蛋白(ELIP)基因在竹子光保护中的作用,为进一步阐述竹子光保护机制提供参考依据。  方法  以毛竹Phyllostachys edulis实生苗为材料,在前期研究的基础上克隆毛竹ELIP基因,利用qRT-PCR技术研究其在不同光照诱导下的表达谱,同时通过在拟南芥Arabidopsis thaliana中异位表达对1个基因的功能进行初步鉴定。  结果  克隆获得了3个毛竹ELIP基因(PeELIP1、PeELIP2和PeELIP3),分别编码165、179和182个氨基酸。蛋白结构分析表明:3个PeELIPs蛋白均具有典型的捕光叶绿素a/b结合蛋白功能域,含3个α-螺旋跨膜结构,属于叶绿素a/b结合蛋白超家族。序列比对及进化分析表明:PeELIPs与水稻Oryza sativa、玉米Zea mays等单子叶植物的ELIPs相似性较高,同源性达72%以上,聚类在同一分支。qRT-PCR分析表明:3个PeELIPs基因在毛竹黄化苗中仅检测到微弱表达,光照处理使3个基因的表达量均显著上调;同时在正常毛竹实生苗叶片中,随着光照强度的增强和强光胁迫处理时间的延长,3个PeELIPs基因的表达量都显著上调。过表达PeELIP3可减缓转基因拟南芥在强光下Fv/Fm的下降幅度,但未影响转基因植株的非光化学猝灭系数。  结论  毛竹中至少存在3个PeELIPs,且其表达均受光照的诱导。过量表达PeELIP3能够减缓转基因拟南芥受光抑制的程度,具有一定的光保护作用。图8表1参40

English Abstract

娄永峰, 高志民. 毛竹早期光诱导蛋白基因克隆及功能分析[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200237
引用本文: 娄永峰, 高志民. 毛竹早期光诱导蛋白基因克隆及功能分析[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20200237
LOU Yongfeng, GAO Zhimin. Cloning and functional analysis of early light induced protein genes of Phyllostachys edulis[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200237
Citation: LOU Yongfeng, GAO Zhimin. Cloning and functional analysis of early light induced protein genes of Phyllostachys edulis[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20200237

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