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Isolation and characterization of a plastidic glutamine synthetase (GS) gene in laticifers of Hevea brasiliensis Muell. Arg.

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Abstract

Glutamine synthetase (GS) in plants is a crucial enzyme for nitrogen assimilation and defending against biotic and abiotic stresses. In the present study, a full-length cDNA, designated HbGS2, was isolated from Hevea brasiliensis by amplification of cDNA ends method. HbGS2 was 1726 bp in length and contained a 1299-bp open reading frame encoding a putative protein of 433 amino acids. The predicted molecular mass of HbGS2 was 47.76 kilodalton with an isoelectric point (pI) of 8.06. Bioinformatics analysis showed that it contained the GS conserved elements and belonged to the plastidic member. Cis-regulatory element analysis showed the presence of stress-responsive elements and plastid-specific elements in the promoter region of HbGS2. Overproduction of recombinant HbGS2 protein gave the Escherichia coli cells or yeast more tolerance to heavy metal and hydrogen peroxide. The transcription level of HbGS2 and the activity of HbGS in Frey-Wyssling particles were significantly repressed by ethylene application. HbGS2 should be located in the Frey-Wyssling particles and may confer these organelles that are homologous to plastids a role in defending against various abiotic stresses.

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Abbreviations

AsA–GSH:

Ascorbate–glutathione

CK:

Control check

EU:

Extension unit

ETH:

Ethylene

GS:

Glutamine synthetase

H2O2 :

Hydrogen peroxide

IPTG:

Isopropyl-beta-d-thiogalactopyranoside

kDa:

Kilodalton

LB:

Luria–Bertani

ORF:

Open reading frame

pI:

Isoelectric point

QRT-PCR:

Quantitative real-time polymerase chain reaction

RACE:

Rapid amplification of cDNA ends

ROS:

Reactive oxygen species

SC:

Synthetic complete medium

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

References

  • Allen RD, Bernier F, Lessard PA, Beachy RN (1989) Nuclear factors interact with a soybean beta-conglycinin enhancer. Plant Cell 1:623–631. doi:10.1105/tpc.1.6.623

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bernard SM, Habash DZ (2009) The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling. New Phytol 182:608–620. doi:10.1111/j.1469-8137.2009.02823.x

    Article  CAS  PubMed  Google Scholar 

  • Boter M, Ruiz-Rivero O, Abdeen A, Prat S (2004) Conserved MYC transcription factors play a key role in jasmonate signaling both in tomato and Arabidopsis. Genes Dev 18:1577–1591. doi:10.1101/gad.297704

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Busk PK, Jensen AB, Pages M (1997) Regulatory elements in vivo in the promoter of the abscisic acid responsive gene rab17 from maize. Plant J 11:1285–1295. doi:10.1046/j.1365-313X.1997.11061285.x

    Article  CAS  PubMed  Google Scholar 

  • Cercos M, Gomez-Cadenas A, Ho THD (1999) Hormonal regulation of a cysteine proteinase gene, EPB-1, in barley aleurone layers: cis- and trans-acting elements involved in the co-ordinated gene expression regulated by gibberellins and abscisic acid. Plant J 19:107–118. doi:10.1046/j.1365-313X.1999.00499.x

    Article  CAS  PubMed  Google Scholar 

  • Chao J, Chen Y, Wu S, Tian WM (2015) Comparative transcriptome analysis of latex from rubber tree clone CATAS8-79 and PR107 reveals new cues for the regulation of latex regeneration and duration of latex flow. BMC Plant Biol 15:104. doi:10.1186/s12870-015-0488-3

    Article  PubMed  PubMed Central  Google Scholar 

  • Cheng HM, Chern Y, Chen IH, Liu C, Li SH, Chun SJ, Rigo F, Bennett CF, Deng N, Feng Y, Lin CS, Yan YT, Cohen SN, Cheng TH (2015) Effects on murine behavior and lifespan of selectively decreasing expression of mutant huntingtin allele by supt4h knockdown. PLoS Genet 11:e1005043. doi:10.1371/journal.pgen.1005043

    Article  PubMed  PubMed Central  Google Scholar 

  • Chinnusamy V, Ohta M, Kanrar S, Lee BH, Hong X, Agarwal M, Zhu JK (2003) ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev 17:1043–1054. doi:10.1101/gad.1077503

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chow KS, Wan KL, Isa MN, Bahari A, Tan SH, Harikrishna K, Yeang HY (2007) Insights into rubber biosynthesis from transcriptome analysis of Hevea brasiliensis latex. J Exp Bot 58:2429–2440. doi:10.1093/jxb/erm093

    Article  CAS  PubMed  Google Scholar 

  • Chow KS, Mat-Isa MN, Bahari A, Ghazali AK, Alias H, Mohd-Zainuddin Z, Hoh CC, Wan KL (2012) Metabolic routes affecting rubber biosynthesis in Hevea brasiliensis latex. J Exp Bot 63:1863–1871. doi:10.1093/jxb/err363

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cyert MS, Philpott CC (2013) Regulation of cation balance in Saccharomyces cerevisiae. Genetics 193:677–713. doi:10.1534/genetics.112.147207

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • d’Auzac J, Jacob JL (1989) The composition of latex from Hevea brasiliensis as a laticiferous cytoplasm. In: d’Auzac J, Jacob JL, Chrestin L (eds) Physiology of rubber tree latex. CRC Press, Boca Raton, pp 59–96

    Google Scholar 

  • Dai L, Kang G, Li Y, Nie Z, Duan C, Zeng R (2013) In-depth proteome analysis of the rubber particle of Hevea brasiliensis (para rubber tree). Plant Mol Biol 82:155–168. doi:10.1007/s11103-013-0047-y

    Article  CAS  PubMed  Google Scholar 

  • Degenhardt J, Tobin EM (1996) A DNA binding activity for one of two closely defined phytochrome regulatory elements in an Lhcb promoter is more abundant in etiolated than in green plants. Plant Cell 8:31–41. doi:10.1105/tpc.8.1.31

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Díaz P, Betti M, Sánchez DH, Udvardi MK, Monza J, Márquez AJ (2010) Deficiency in plastidic glutamine synthetase alters proline metabolism and transcriptomic response in Lotus japonicus under drought stress. New Phytol 188:1001–1013. doi:10.1111/j.1469-8137.2010.03440.x

    Article  PubMed  Google Scholar 

  • El-Maarouf-Bouteau H, Sajjad Y, Bazin J, Langlade N, Cristescu SM, Balzergue S, Baudouin E, Bailly C (2015) Reactive oxygen species, abscisic acid and ethylene interact to regulate sunflower seed germination. Plant Cell Environ 38:364–374. doi:10.1111/pce.12371

    Article  CAS  PubMed  Google Scholar 

  • Eschbach JM, Lacrotte R (1989) Factors influencing response to hormonal yield stimulation: limits of this stimulation. In: d’Auzac J, Jacob J-L, Chrestin H (eds) Physiology of rubber tree latex. CRC Press, Boca Raton, pp 321–342

    Google Scholar 

  • Eulgem T, Rushton PJ, Robatzek S, Somssich IE (2000) The WRKY superfamily of plant transcription factors. Trends Plant Sci 5:199–206. doi:10.1016/S1360-1385(00)01600-9

    Article  CAS  PubMed  Google Scholar 

  • Feyder S, De Craene JO, Bär S, Bertazzi DL, Friant S (2015) Membrane trafficking in the yeast Saccharomyces cerevisiae model. Int J Mol Sci 16:1509–1525. doi:10.3390/ijms16011509

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fusada N, Masuda T, Kuroda H, Shimada H, Ohta H, Takamiya K (2005) Identification of a novel cis-element exhibiting cytokinin-dependent protein binding in vitro in the 5′-region of NADPH-protochlorophyllide oxidoreductase gene in cucumber. Plant Mol Biol 59:631–645. doi:10.1007/s11103-005-0579-x

    Article  CAS  PubMed  Google Scholar 

  • Guo Y, Cai Z, Gan S (2004) Transcriptome of Arabidopsis leaf senescence. Plant Cell Environ 27:521–549

    Article  CAS  Google Scholar 

  • Hao BZ, Wu JL (2000) Laticifer differentiation in Hevea brasiliensis: induction by exogenous jasmonic acid and linolenic acid. Ann Bot 85:37–43

    Article  CAS  Google Scholar 

  • Hoffman BM, Lukoyanov D, Yang ZY, Dean DR, Seefeldt LC (2014) Mechanism of nitrogen fixation by nitrogenase: the next stage. Chem Rev 114:4041–4062. doi:10.1021/cr400641x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hudson ME, Quail PH (2003) Identification of promoter motifs involved in the network of phytochrome A-regulated gene expression by combined analysis of genomic sequence and microarray data. Plant Physiol 133:1605–1616. doi:10.1104/pp.103.030437

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kapoor S, Sugiura M (1999) Identification of two essential sequence elements in the nonconsensus type II PatpB-290 plastid promoter by using plastid transcription extracts from cultured tobacco BY-2 cells. Plant Cell 11:1799–1810. doi:10.1105/tpc.11.9.1799

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Koyama T (2014) The roles of ethylene and transcription factors in the regulation of onset of leaf senescence. Front Plant Sci 25:650. doi:10.3389/fpls.2014.00650

    Google Scholar 

  • Lagrange T, Franzetti B, Axelos M, Mache R, Lerbs-Mache S (1993) Structure and expression of the nuclear gene encoding for the chloroplast ribosomal protein L21: developmental regulation of a house-keeping gene by alternative promoters. Mol Cell Biol 13:2614–2622. doi:10.1128/MCB.13.4.2614

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li D, Deng Z, Liu C, Zhao M, Guo H, Xia Z, Liu H (2014) Molecular cloning, expression profiles, and characterization of a novel polyphenol oxidase (PPO) gene in Hevea brasiliensis. Biosci Biotechnol Biochem 78:1648–1655. doi:10.1080/09168451

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Chen YY, Yang SG, Tian WM (2015) Cloning and characterization ofHbMT2a, a metallothionein gene from Hevea brasiliensis Muell. Arg differently responds to abiotic stress and heavy metals. Biochem Biophys Res Commun 461:95–101. doi:10.1016/j.bbrc.2015.03.175

    Article  CAS  PubMed  Google Scholar 

  • Ma N, Cai L, Lu W, Tan H, Gao J (2005) Exogenous ethylene influences flower opening of cut roses (Rosa hybrida) by regulating the genes encoding ethylene biosynthesis enzymes. Sci China C Life Sci 48:434–444

    Article  CAS  PubMed  Google Scholar 

  • Maeda K, Kimura S, Demura T, Takeda J, Ozeki Y (2005) DcMYB1 acts as a transcriptional activator of the carrot phenylalanine ammonia-lyase gene (DcPAL1) in response to elicitor treatment, UV-B irradiation and the dilution effect. Plant Mol Biol 59:739–775. doi:10.1007/s11103-005-0910-6

    Article  CAS  PubMed  Google Scholar 

  • Manzara T, Gruissem W (1988) Organization and expression of the genes encoding ribulose-1,5-bisphosphate carboxylase in higher plants. Photosynth Res 16:117–139. doi:10.1007/BF00039489

    Article  CAS  PubMed  Google Scholar 

  • Martin A, Lee J, Kichey T (2006) Two cytosolic glutamine synthetase isoforms of maize are specifically involved in the control of grain production. Plant Cell 18:3252–3274. doi:10.1105/tpc.106.042689

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Masclaux-Daubresse C, Daniel-Vedele F, Dechorgnat J, Chardon F, Gaufichon L, Suzuki A (2010) Nitrogen uptake, assimilation and remobilization in plants: challenges for sustainable and productive agriculture. Ann Bot 105:1141–1157. doi:10.1093/aob/mcq028

    Article  PubMed  PubMed Central  Google Scholar 

  • Mathis DJ, Chambon P (1981) The SV40 early region TATA box is required for accurate in vitro initiation of transcription. Nature 290:310–315

    Article  CAS  PubMed  Google Scholar 

  • Miflin BJ, Habash DZ (2002) The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops. J Exp Bot 53:979–987

    Article  CAS  PubMed  Google Scholar 

  • Morita A, Umemura T, Kuroyanagi M, Futsuhara Y, Perata P, Yamaguchi J (1998) Functional dissection of a sugar-repressed alpha-amylase gene (Ramy1A) promoter in rice embryos. FEBS Lett 423:81–85. doi:10.1016/S0014-5793(98)00067-2

    Article  CAS  PubMed  Google Scholar 

  • Orozco-Cardenas ML, Ryan C (1999) Hydrogen peroxide is generated systemically in plant leaves by wounding and systemin via the octadecanoid pathway. Proc Natl Acad Sci USA 96:6553–6557. doi:10.1073/pnas.96.11.6553

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pageau K, Reisdorf-Cren M, Morot-Gaudry JF, Masclaux-Daubresse C (2006) The two senescence-related markers, GS1 (cytosolic glutamine synthetase) and GDH (glutamate dehydrogenase), involved in nitrogen mobilization, are differentially regulated during pathogen attack and by stress hormones and reactive oxygen species in Nicotiana tabacum L leaves. J Exp Bot 57:547–557. doi:10.1093/jxb/erj035

    Article  CAS  PubMed  Google Scholar 

  • Pan Y, Wang YC, Zhang DW, Yang CP (2010) Cloning and stress tolerance analysis of an LbGRP gene in Limonium bicolor. Yi Chuan 32:278–86

    Article  CAS  PubMed  Google Scholar 

  • Pognani M, Barrena R, Font X, Sánchez A (2012) A complete mass balance of a particle, combined anaerobic/aerobic municipal source-separated waste treatment plant. Waste Manag. doi:10.1016/j.wasman.2011.12.018

    PubMed  Google Scholar 

  • Premarkumari D, Panikkar AON (1989) Anatomy and ultracytology of latex vessels. In: d’Auzac J, Jacob J-L, Chrestin H (eds) Physiology of rubber tree latex. CRC Press, Boca Raton, pp 67–87

    Google Scholar 

  • Pucciariello C, Parlanti S, Banti V, Novi G, Perata P (2012) Reactive oxygen species-driven transcription in Arabidopsis under oxygen deprivation. Plant Physiol 159:184–196. doi:10.1104/pp.111.191122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pujade-Renaud V, Clement A, Perrot-Rechenmann C, Prevot JC, Chrestin H, Jacob JL, Guern J (1994) Ethylene-induced increase in glutamine synthetase activity and mRNA Levels in Hevea brasiliensis latex cells. Plant Physiol 105:127–132. doi:10.1104/pp.105.1.127

    CAS  PubMed  PubMed Central  Google Scholar 

  • Rieping M, Schoffl F (1992) Synergistic effect of upstream sequences, CCAAT box elements, and HSE sequences for enhanced expression of chimaeric heat shock genes in transgenic tobacco. Mol Gen Genet 231:226–232

    CAS  PubMed  Google Scholar 

  • Ross EJ, Stone JM, Elowsky CG, Arredondo-Peter R, Klucas RV, Sarath G (2004) Activation of the Oryza sativa non-symbiotic haemoglobin-2 promoter by the cytokinin-regulated transcription factor, ARR1. J Exp Bot 55:1721–1731. doi:10.1093/jxb/erh211

    Article  CAS  PubMed  Google Scholar 

  • Savatin DV, Gramegna G, Modesti V, Cervone F (2014) Wounding in the plant tissue: the defense of a dangerous passage. Front Plant Sci 16:470. doi:10.3389/fpls.2014.00470

    Google Scholar 

  • Shirsat A, Wilford N, Croy R, Boulter D (1989) Sequences responsible for the tissue specific promoter activity of a pea legumin gene in tobacco. Mol Gen Genet 215:326–331

    Article  CAS  PubMed  Google Scholar 

  • Silva L, Carvalho H (2013) Possible role of glutamine synthetase in the NO signaling response in root nodules by contributing to the antioxidant defenses. Front Plant Sci 4:372. doi:10.3389/fpls.2013.00372

    Article  PubMed  PubMed Central  Google Scholar 

  • Simpson SD, Nakashima K, Narusaka Y, Seki M, Shinozaki K, Simpson SD, Nakashima K, Narusaka Y, Seki M, Shinozaki K (2003) Two different novel cis-acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark-induced senescence. Plant J 33:259–270. doi:10.1046/j.1365-313X.2003.01624.x

    Article  CAS  PubMed  Google Scholar 

  • Spence C, Alff E, Johnson C, Ramos C, Donofrio N, Sundaresan V, Bais H (2014) Natural rice rhizospheric microbes suppress rice blast infections. BMC Plant Biol 14:130. doi:10.1186/1471-2229-14-130

    Article  PubMed  PubMed Central  Google Scholar 

  • Steffens B (2014) The role of ethylene and ROS in salinity, heavy metal, and flooding responses in rice. Front Plant Sci 5:685. doi:10.3389/fpls.2014.00685

    Article  PubMed  PubMed Central  Google Scholar 

  • Tang C, Huang D, Yang J, Liu S, Sakr S, Li H, Zhou Y, Qin Y (2010) The sucrose transporter HbSUT3 plays an active role in sucrose loading to laticifer and rubber productivity in exploited trees of Hevea brasiliensis (para rubber tree). Plant Cell Environ 33:1708–1720. doi:10.1111/j.1365-3040.2010.02175.x

    Article  CAS  PubMed  Google Scholar 

  • Thomsen HC, Eriksson D, Møller IS, Schjoerring JK (2014) Cytosolic glutamine synthetase: a target for improvement of crop nitrogen use efficiency. Trends Plant Sci 19:656–663. doi:10.1016/j.tplants.2014.06.002

    Article  CAS  PubMed  Google Scholar 

  • Ulmasov T, Hagen G, Guilfoyle TJ (1999) Dimerization and DNA binding of auxin response factors. Plant J 19:309–319. doi:10.1046/j.1365-313X.1999.00538.x

    Article  CAS  PubMed  Google Scholar 

  • Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K (1993) An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant Cell 5:1529–1539. doi:10.1105/tpc.5.11.1529

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vieweg MF, Fruhling M, Quandt HJ, Heim U, Baumlein H, Puhler A, Kuster H, Andreas MP (2004) The promoter of the Vicia faba L. leghemoglobin gene VfLb29 is specifically activated in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots from different legume and nonlegume plants. Mol Plant Microbe Interact 17:62–69. doi:10.1094/MPMI.2004.17.1.62

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Shi M, Wang D, Chen Y, Cai F, Zhang S, Wang L, Tong Z, Tian WM (2013) Comparative proteomics of primary and secondary lutoids reveals that chitinase and glucanase play a crucial combined role in rubber particle aggregation in Hevea brasiliensis. J Proteome Res 12:5146–5159. doi:10.1021/pr400378c

    Article  CAS  PubMed  Google Scholar 

  • Xia XJ, Zhou YH, Shi K, Zhou J, Foyer CH, Yu JQ (2015) Interplay between reactive oxygen species and hormones in the control of plant development and stress tolerance. J Exp Bot 66:2839–2856

    Article  CAS  PubMed  Google Scholar 

  • Yanagisawa S, Schmidt RJ (1999) Diversity and similarity among recognition sequences of Dof transcription factors. Plant J 17:209–214. doi:10.1046/j.1365-313X.1999.00363.x

    Article  CAS  PubMed  Google Scholar 

  • Yang ZP, Li HL, Guo D, Tian WM, Peng SQ (2012) Molecular characterization of a novel 14-3-3 protein gene (Hb14-3-3c) from Hevea brasiliensis. Mol Biol Rep 39:4491–4497. doi:10.1007/s11033-011-1239-7

    Article  CAS  PubMed  Google Scholar 

  • Yip WK, Yang SF (1988) Cyanide metabolism in relation to ethylene production in plant tissues. Plant Physiol 88:473–476

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou DX (1999) Regulatory mechanism of plant gene transcription by Binding of GT-1-like factors and GT-factors. Trends Plant Sci 4:210–214

    Article  PubMed  Google Scholar 

  • Zhu J, Zhang Z (2009) Ethylene stimulation of latex production in Hevea brasiliensis. Plant Signal Behav 4:1072–1074

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31170642) and the Earmarked Fund for Modern Agro-industry Technology Research System (CARS-34-GW1).

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  1. Ministry of Agriculture Key Laboratory of Biology and Genetic Resources of Rubber Tree/State Key Laboratory Breeding Base of Cultivation and Physiology for Tropical Crops, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, 571737, Hainan, People’s Republic of China

    Jinquan Chao, Shixin Zhang, Yueyi Chen & Wei-Min Tian

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  1. Jinquan Chao
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Chao, J., Zhang, S., Chen, Y. et al. Isolation and characterization of a plastidic glutamine synthetase (GS) gene in laticifers of Hevea brasiliensis Muell. Arg.. Acta Physiol Plant 38, 90 (2016). https://doi.org/10.1007/s11738-016-2107-9

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