中国农业科学 ›› 2019, Vol. 52 ›› Issue (19): 3404-3416.doi: 10.3864/j.issn.0578-1752.2019.19.011
收稿日期:
2019-05-05
接受日期:
2019-07-12
出版日期:
2019-10-01
发布日期:
2019-10-11
通讯作者:
谢让金
作者简介:
葛廷,E-mail:m13364018674@163.com。
基金资助:
GE Ting,HUANG Xue,XIE RangJin()
Received:
2019-05-05
Accepted:
2019-07-12
Online:
2019-10-01
Published:
2019-10-11
Contact:
RangJin XIE
摘要:
【目的】 多聚半乳糖醛酸酶是一类参与细胞壁降解的水解酶,在植物生长发育和器官脱落过程中发挥着重要作用。本研究克隆柑橘CitPG34及其启动子(CitPG34-P)并进行表达分析,为深入研究柑橘PG在幼果脱落过程的生物功能奠定基础。【方法】 以‘塔罗科’血橙(Citrus sinensis L. Osbeck)为材料,克隆CitPG34及其启动子,利用ProtParam、Cello、CLUSTALX、MEGA5.2、PlantCARE等软件对其蛋白特性及启动子顺式作用元件进行分析预测;利用实时荧光定量PCR(qRT-PCR)分析CitPG34在不同组织以及柑橘幼果脱落过程中的表达水平。采用同源重组的方法构建pCAMBIA1302-CitPG34-GFP融合蛋白表达载体和CitPG34启动子表达载体(CitPG34-P::gus),分别用于亚细胞定位和启动子活性分析。【结果】 从‘塔罗科’血橙幼果离层中克隆获得CitPG34,其ORF为1 194 bp,编码397个氨基酸,预测蛋白分子量为41.47 kD,理论等电点为5.19,其不稳定系数为30.23,表明CitPG34属于稳定蛋白;通过在线软件TMHMM分析发现:CitPG34为跨膜蛋白,具有一个跨膜结构,位于第7—29位氨基酸之间。在CitPG34二级结构中,α-螺旋结构约占15.37%,扩展链约占29.72%,无规则卷曲约占54.91%,与其三级结构预测基本一致。NJ树分析显示CitPG34与西洋梨PcPG3(BAF42034)亲缘关系最近,表明CitPG34可能与果实脱落和软化相关。qRT-PCR分析表明,CitPG34在花中表达量最高,在根、叶、离层A、离层C中表达量较低,在幼果中几乎不表达。1-氨基环丙烷羧酸(ACC)处理果梗后能显著提高离层A中CitPG34的表达水平,相反IAA抑制其转录。此外,在柑橘幼果正常脱落过程中,CitPG34表达明显升高。亚细胞定位发现,CitPG34主要位于细胞壁。克隆获取CitPG34起始密码子(ATG)前2 075 bp启动子序列(CitPG34-P),PlantCare预测发现,在CitPG34-P序列上存在多种顺式调控元件,如核心启动元件TATA-box、增强子元件CAAT-box以及脱落酸响应元件ABRE等。将CitPG34-P::gus转入烟草,通过GUS组织化学染色发现,该启动子受乙烯诱导,主要在叶脉和毛状体中表达。【结论】 CitPG34的ORF长度为1 194 bp,可编码397个氨基酸,其蛋白主要位于细胞壁;该基因具有明显的组织特异性,在花中表达最高;CitPG34表达量与柑橘幼果脱落显著相关。上述结果表明,CitPG34在柑橘幼果脱落和花发育过程中可能发挥着重要的生物功能。
葛廷,黄雪,谢让金. 柑橘CitPG34的克隆、定位与表达分析[J]. 中国农业科学, 2019, 52(19): 3404-3416.
GE Ting,HUANG Xue,XIE RangJin. Cloning, Subcellular Localization and Expression Analysis of CitPG34 in Citrus[J]. Scientia Agricultura Sinica, 2019, 52(19): 3404-3416.
表1
本试验所用引物"
引物名称 Primers name | 引物序列 Sequence of primers (5′→3′) | 目的 Purpose |
---|---|---|
OE-CitPG34-F | GCTTTGAAACCATGTCAACATC | 基因全长克隆 Full-length gene cloning |
OE-CitPG34-R | TAGGAACTGCCTCTAAGTAAGC | |
CitPG34-F | ATTAGCATCCGTAGATAGCC | 实时荧光定量 Real-time PCR |
CitPG34-R | CACAGTCATCACCAGTCCC | |
P- CitPG34-F | TGATTACGCCAAGCTTACCGTCTATGCTACTGATGTGA | 启动子克隆 Promoter cloning |
P- CitPG34-R | AGGGACTGACCACCCTGATGTTGACATGGTTTCAAAGC | |
Cit-Actin-F | CCCCATCGTTACCGTCCAG | 内参基因 Actin gene |
Cit-Actin-R | CGCCTTGCCAGTTGAATATCC | |
Sub-CitPG34-F | GGGGGACTCTTGACCATGTCAACATCAATGTCTTTGCC | 亚细胞定位 Subcellular localization |
sub-CitPG34-R | CTAGTCAGATCTACCATGACACCAGAACCACCACCAGA ACCAGTCTTCAAGCAACTATTTGGAAG | |
pCAMBIA1302-F | CCTTCACCCTCTCCACTGACAG | 亚细胞定位 Subcellular localization |
pCAMBIA1302-R | CCTTCGCAAGACCCTTCCTCTA |
图3
CitPG34与其他物种PG蛋白的氨基酸系列比对 AtPG1:拟南芥Arabidopsis thaliana,NP191310;AtPG2:拟南芥Arabidopsis thaliana,NP850359;AdPG:猕猴桃Actinidia deliciosa,L12019;BrnPG:欧洲油菜Brassica napus,NP001302495;CmPG1:甜瓜Cucumis melo,AF062465;CmPG2:甜瓜Cucumis melo,AF062466;CpPG4:番木瓜Carica papaya,GQ479796;DkPG3:柿Diospyros kaki,EU816199;LcPG:荔枝Litchi chinensis,AFW04075;GmPG6:大豆Glycine max,ABD62085;MaPG3:小果野蕉Musa acuminata,AY603339;NtPG1:烟草Nicotiana tabacum,AHG12641;MdPG:苹果Malus domestica,L27743;PaPG:杏Prunus armeniaca,ADT82706;PcPG:西洋梨Pyrus communis,CAH18935;PcPG1:西洋梨Pyrus communis,AB084461;PcPG3:西洋梨Pyrus communis,BAF42034;PdPG1:欧洲李Prunus domestica,DQ375247;PpPG:碧桃Prunus persica,EF568784;PpPG1:碧桃Prunus persica,BAH56488;PpPG2:碧桃Prunus persica,CAA54448;TAPG1:番茄Solanum lycopersicum,NM001246866;TAPG2:番茄Solanum lycopersicum,U70480;TAPG4:番茄Solanum lycopersicum,U70481;VvPG1:葡萄Vitis vinifera,AY043233;VvPG2:葡萄Vitis vinifera,EU078975;ZmPG:玉米Zea mays,P26216 I:SPNTDG结构域;II:GDDC结构域;III:CGPGHG结构域;IV:RIK结构域"
表2
CitPG34启动子序列主要顺式作用元件"
顺式作用元件 cis-element | 序列 Sequence | 功能 Function | 数量 Number |
---|---|---|---|
ABRE | CACGTG; ACGTG; AACCCGG | 脱落酸应答相关的顺式作用元件 Cis-acting element involved in the abscisic acid responsiveness | 7 |
ARE | AAACCA | 厌氧诱导必须顺式作用元件 Cis-acting regulatory element essential for the anaerobic induction | 2 |
CGTCA-motif | CGTCA | 甲基茉莉酸应答相关的顺式作用元件 Cis-acting regulatory element involved in the MeJA-responsiveness | 2 |
MBS | CAACTG | 参与干旱诱导的MYB结合位点 MYB binding site involved in drought-inducibility | 3 |
MBSI | aaaAaaC(G/C)GTTA | 类黄酮生物合成基因调控相关的MYB结合位点 MYB binding site involved in flavonoid biosynthetic genes regulation | 1 |
MYB | TAACCA | 赤霉素作用元件 Gibberellin action element | 3 |
MYB-like sequence | TAACCA | 赤霉素作用元件 Gibberellin action element | 3 |
MYC | CATTTG; CATGTG | MYC结合位点 MYC binding site | 2 |
TC-rich repeats | ATTCTCTAAC | 胁迫响应元件 Cis-acting element involved in defense and stress responsiveness | 1 |
TCA-element | CCATCTTTTT | 水杨酸应答元件 Cis-acting element involved in salicylic acid responsiveness | 1 |
TGACG-motif | TGACG | 茉莉酸甲酯调控元件 Cis-acting regulatory element involved in the MeJA-responsiveness | 2 |
WUN-motif | AAATTTCCT | 机械伤应答元件 wound-responsive element | 1 |
as-1 | TGACG | 茉莉酸甲酯调控元件 Cis-acting regulatory element involved in the MeJA-responsiveness | 2 |
Box-W1 | TTGACC | 真菌诱导应答元件 Fungal induced response elements | 1 |
[1] | 刘志良 . 柑橘过量落花落果的原因及防止对策. 中国农技推广, 2013,29(9):28-30. |
LIU Z L . Reasons for excessive flowering and fruit dropping of citrus and preventive measures. China Agricultural Technology Extension, 2013,29(9):28-30. (in Chinese) | |
[2] |
潘小婷, 张静, 葛廷, 马岩岩, 邓烈, 何绍兰, 易时来, 郑永强, 吕强, 谢让金 . 柑橘CitCEP基因家族的鉴定及对逆境和激素的响应. 中国农业科学, 2018,51(16):3147-3158.
doi: 10.3864/j.issn.0578-1752.2018.16.010 |
PAN X T, ZHANG J, GE T, MA Y Y, DENG L, HE S L, YI S L, ZHENG Y Q, LÜ Q, XIE R J . Identification of citrus CitCEP genes and their transcriptional response to stress and hormone treatments. Scientia Agricultura Sinica, 2018,51(16):3147-3158. (in Chinese)
doi: 10.3864/j.issn.0578-1752.2018.16.010 |
|
[3] | RIOV J . A polygalacturonase from citrus leaf explants: Role in abscission. Plant Physiology, 1974,53(2):312-316. |
[4] | NAKANO T, ITO Y . Molecular mechanisms controlling plant organ abscission. Plant Biotechnology, 2013,30(3):209-216. |
[5] | CARMEN RODRÍGUEZ-GACIO M D C, NICOLÁS C, MATILLA A J . Cloning and analysis of a cDNA encoding an endo-polygalacturonase expressed during the desiccation period of the silique-valves of turnip-tops (Brassica rapa L. cv. Rapa). Journal of Plant Physiology, 2004,161(2):219-227. |
[6] | QUESADA M, BLANCO-PORTALES R, POSÉ S, GARCÍA-GAGO J A, JIMÉNEZ-BERMÚDEZ S, MUN˜OZ-SERRANO A, CABALLERO J L, PLIEGO-ALFARO F, MERCADO J A, MUÑOZ-BLANCO J . Antisense down-regulation of the FaPG1 gene reveals an unexpected central role for polygalacturonase in strawberry fruit softening. Plant Physiology, 2009,150(2):1022-1032. |
[7] | POSE S, PANIAGUA C, CIFUENTES M, BLANCO-PORTALES R, QUWSADA M A, MERCADO J A . Insights into the effects of polygalacturonase FaPG1 gene silencing on pectin matrix disassembly, enhanced tissue integrity, and firmness in ripe strawberry fruits. Journal of Experimental Botany, 2013,64(12):3803-3815. |
[8] | GONZÁLEZ-CARRANZA Z H, ELLIOTT K A, ROBERTS J A . Expression of polygalacturonases and evidence to support their role during cell separation processes in Arabidopsis thaliana. Journal of Experimental Botany, 2007,58(13):3719-3730. |
[9] | LYU M L, LIANG Y, YU Y J, MA Z M, SONG L M, YUE X Y, CAO J S . Identification and expression analysis of BoMF25, a novel polygalacturonase gene involved in pollen development of Brassica oleracea. Plant Reproduction, 2015,28(2):121-132. |
[10] | ATKINSON R G, SCHRODER R, HALLETT I C, COHEN D, MACRAE E A . Overexpression of polygalacturonase in transgenic apple trees leads to a range of novel phenotypes involving changes in cell adhesion. Plant Physiology, 2002,129(1):122-133. |
[11] | ROONGSATTHAM P, MORCILLO F, JANTASURIYARAT C, PIZOT M, MOUSSU S, JAYAWEERA D, COLLIN M, GONZÁLEZ- CARRANZA Z H, AMBLARD P, TREGEAR J W, TRAGOONRUNG S, VERDEIL J, TRANBARGER T J . Temporal and spatial expression of polygalacturonase gene family members reveals divergent regulation during fleshy fruit ripening and abscission in the monocot species oil palm. BMC Plant Biology, 2012,12(1):150. |
[12] | JIANG C Z, LU F, IMSABAI W, MEIR S, REID M S . Silencing polygalacturonase expression inhibits tomato petiole abscission. Journal of Experimental Botany, 2008,59(4):973. |
[13] | PENG G, WU J Y, LU W J, LI J G . A polygalacturonase gene clustered into clade E involved in lychee fruitlet abscission. Scientia Horticulturae, 2013,150:244-250. |
[14] | RASCIO N, CASADORO G, RAMINA A, MASIA A . Structural and biochemical aspects of peach fruit abscission (Prunus persica L. Batsch). Planta, 1985,164(1):1-11. |
[15] | TAYLOR J E, WEBB S T J, COUPE S A, TUCKER G A, ROBERTS J A . Changes in polygalacturonase activity and solubility of polyuronides during ethylene-stimulated leaf abscission in Sambucus nigra. Journal of Experimental Botany, 1993,44(1):93-98. |
[16] | KALAITZIS P, TUCKER S M L . Three different polygalacturonases are expressed in tomato leaf and flower abscission, each with a different temporal expression pattern. Plant Physiology, 1997,113(4):1303-1308. |
[17] | CHENG C Z, ZHANG L Y, YANG X L, ZHONG G Y . Profiling gene expression in citrus fruit calyx abscission zone (AZ-C) treated with ethylene. Molecular Genetics and Genomics, 2015,290(5):1991-2006. |
[18] | MERELO P, AGUSTÍJ, ARBONA V, COSTA M L, ESTORNELL L H, GÓMEZ-CADENAS A, COIMBRA S, GÓMEZ M D, PÉREZ-AMADOR M A, DOMINGO C, TALÓN M, TADEO F R . Cell wall remodeling in abscission zone cells during ethylene- promoted fruit abscission in citrus. Frontiers in Plant Science, 2017,8:126. |
[19] | XIE R J, GE T, ZHANG J, PAN X T, MA Y Y, YI S L, ZHENG Y Q . The molecular events of IAA inhibiting citrus fruitlet abscission revealed by digital gene expression profiling. Plant Physiology and Biochemistry, 2018,130:192-204. |
[20] | GONZÁLEZ-CARRANZA Z H, WHITELAW C A, SWARUP R, ROBERTS J A . Temporal and spatial expression of a polygalacturonase during leaf and flower abscission in oilseed rape and Arabidopsis. Plant Physiology, 2002,128(2):534-543. |
[21] | MARKOVIC O, JANECEK S . Pectin degrading glycoside hydrolases of family 28: Sequence structural features, specificities and evolution. Protein Engineering Design and Selection, 2001,14(9):615-631. |
[22] | SITRIT Y, HADFIELD K A, BENNETT A B, BRADFORD K J, DOWNIE A B . Expression of a polygalacturonase associated with tomato seed germination. Plant Physiology, 1999,121(2):419-428. |
[23] | SANDER L, CHILD R, ULVSKOV P, ALBRECHTSEN M, BORKHARDT B . Analysis of a dehiscence zone endo-polygalacturonase in oilseed rape (Brassica napus) and Arabidopsis thaliana: Evidence for roles in cell separation in dehiscence and abscission zones, and in stylar tissues during pollen tube growth. Plant Molecular Biology, 2001,46(4):469-479. |
[24] | ROBERTS J A, ELLIOTT K A, GONZÁLEZ-CARRANZA Z H . Abscission, dehiscence, and other cell separation processes. Annual Review of Plant Biology, 2001,53(1):131-158. |
[25] | CHEN H F, SHAO H X, FAN S, MA J J, ZHANG D, HAN M Y . Identification and phylogenetic analysis of the POLYGALACTURONASE gene family in apple. Horticultural Plant Journal, 2016,2(5):241-252. |
[26] | MING Q, YIKE Z, YAN X Y, HAN M Y . identification and expression analysis of polygalacturonase family members during peach fruit softening. International Journal of Molecular Sciences, 2016,17(11):1933. |
[27] | KE X B, WANG H S, LI Y, ZHU B, ZANG Y X, HE Y, CAO J S, ZHU Z J, YU Y J . Genome-wide identification and analysis of polygalacturonase genes in Solanum lycopersicum. International Journal of Molecular Sciences, 2018,19(8):2290. |
[28] | OGAWA M, KAY P SWAIN W S M, . ARABIDOPSIS DEHISCENCE ZONE POLYGALACTURONASE1 (ADPG1), ADPG2, and QUARTET2 are polygalacturonases required for cell separation during reproductive development in Arabidopsis. The Plant Cell, 2009,21(1):216-233. |
[29] | KIM J, SHIU S H, THOMA S, LI W H, PATTERSON S E . Patterns of expansion and expression divergence in the plant polygalacturonase gene family. Genome Biology, 2006,7(9):R87. |
[30] | WANG F F, SUN X, SHI X Y, ZHAI H, TIAN C G, KONG F J, LIU B H, YUAN X H . A global analysis of the polygalacturonase gene family in soybean (Glycine max). PLoS ONE, 2016,11(9):e0163012. |
[31] | YANG Z L, LIU H J, WANG X R, ZENG Q Y . Molecular evolution and expression divergence of the populus polygalacturonase supergene family shed light on the evolution of increasingly complex organs in plants. New Phytologist, 2013,197(4):1353-1365. |
[32] | FABI J P, BROETTO S G, DA SILVA S L G L,, ZHONG S L,, LAJOLO F M,, DO NASCIMENTO J R O, . Analysis of papaya cell wall-related genes during fruit ripening indicates a central role of polygalacturonases during pulp softening. PLoS ONE, 2014,9(8):e105685. |
[33] | DEYTIEUX-BELLEAU C, AMÉLIE V, BERNARD D, GENY L . Pectin methylesterase and polygalacturonase in the developing grape skin. Plant Physiology and Biochemistry, 2008,46(7):638-646. |
[34] | HADFIELD K A, BENNETT A B . Polygalacturonases: Many genes in search of a function. Plant Physiology, 1998,117(2):337-343. |
[35] | BONGHI C, RASCIO N, RAMINA A, CASADORO G . Cellulase and polygalacturonase involvement in the abscission of leaf and fruit explants of peach. Plant Molecular Biology, 1992,20:839-848. |
[36] | HONG S B, SEXTON R, TUCKER M L . Analysis of gene promoters for two tomato polygalacturonases expressed in abscission zones and the stigma. Plant Physiology, 2000,123(3):869-881. |
[37] | LYU M L, YU Y J, JIANG J J, SONG L M, LIANG Y, MA Z M, XIONG X P, CAO J S . BcMF26a and BcMF26b are duplicated polygalacturonase genes with divergent expression patterns and functions in pollen development and pollen tube formation in Brassica campestris. PLoS ONE, 2015,10(7):e0131173. |
[38] | RUI Y, XIAO C W, YI J, KANDEMIR B, WANG J Z, PURI V M, ANDERSON C T . POLYGALACTURONASE INVOLVED IN EXPANSION3 functions in seedling development, rosette growth, and stomatal dynamics in Arabidopsis thaliana. Plant Cell, 2017,29(10):2413. |
[39] | XIAO C, SOMERVILLE C, ANDERSON C T . POLYGALACTURONASE INVOLVED IN EXPANSION1 functions in cell elongation and flower development in Arabidopsis. The Plant Cell, 2014,26(3):1018-1035. |
[40] | KÜHN N, SERRANO A, ABELLO C, ARCE A, ESPINOZA C, GOUTHU S, DELUC L, JOHNSON P A . Regulation of polar auxin transport in grapevine fruitlets (Vitis vinifera L.) and the proposed role of auxin homeostasis during fruit abscission. BMC Plant Biology, 2016,16(1):234. |
[41] | YUAN R C, WU Z C, KOSTENYUK I A, BURNS J K . G- protein-coupled alpha2A-adrenoreceptor agonists differentially alter citrus leaf and fruit abscission by affecting expression of ACC synthase and ACC oxidase. Journal of Experimental Botany, 2005,56(417):1867. |
[42] | VAN DOORN W G, STEAD A D . Abscission of flowers and floral parts. Journal of Experimental Botany, 1997,48(4):821-837. |
[43] | MEIR S, PHILOSOPH-HADAS S, SUNDARESAN S, SELVARAJ K S, BURD S, OPHIR R, KOCHANEK B, REID M S, JIANG C Z, LERS A . Microarray analysis of the abscission-related transcriptome in the tomato flower abscission zone in response to auxin depletion. Plant Signaling & Behavior, 2011,154(4):1929-1956. |
[44] | LI J G, YUAN R C . NAA and ethylene regulate expression of genes related to ethylene biosynthesis, perception, and cell wall degradation during fruit abscission and ripening in ‘Delicious’ apples. Journal of Plant Growth Regulation, 2008,27(3):283-295. |
[45] | ZHU H, DARDICK C D, BEERS E P, CALLANHAN A M, XIA R, YUAN R C . Transcriptomics of shading-induced and NAA-induced abscission in apple (Malus domestica) reveals a shared pathway involving reduced photosynthesis, alterations in carbohydrate transport and signaling and hormone crosstalk. BMC Plant Biology, 2011,11(1):138-138. |
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