Effect of ACGT motif in spatiotemporal regulation of AtAVT6D, which improves tolerance to osmotic stress and nitrogen-starvation.
Arabidopsis thaliana
AtAVT6D
Microprojectile bombardment
Promoter analysis
Xenopus oocyte
Yeast complementation
Journal
Plant molecular biology
ISSN: 1573-5028
Titre abrégé: Plant Mol Biol
Pays: Netherlands
ID NLM: 9106343
Informations de publication
Date de publication:
May 2022
May 2022
Historique:
received:
27
11
2021
accepted:
01
03
2022
pubmed:
5
4
2022
medline:
10
5
2022
entrez:
4
4
2022
Statut:
ppublish
Résumé
Plasma membrane-localized AtAVT6D importing aspartic acid can be targeted to develop plants with enhanced osmotic and nitrogen-starvation tolerance. AtAVT6D promoter can be exploited as a stress-inducible promoter for genetic improvements to raise stress-resilient crops. The AtAVT6 family of amino acid transporters in Arabidopsis thaliana has been predicted to export amino acids like aspartate and glutamate. However, the functional characterization of these amino acid transporters in plants remains unexplored. The present study investigates the expression patterns of AtAVT6 genes in different tissues and under various abiotic stress conditions using quantitative Real-time PCR. The expression analysis demonstrated that the member AtAVT6D was significantly induced in response to phytohormone ABA and stresses like osmotic and drought. The tissue-specific expression analysis showed that AtAVT6D was strongly expressed in the siliques. Taking together these results, we can speculate that AtAVT6D might play a vital role in silique development and abiotic stress tolerance. Further, subcellular localization study showed AtAVT6D was localized to the plasma membrane. The heterologous expression of AtAVT6D in yeast cells conferred significant tolerance to nitrogen-deficient and osmotic stress conditions. The Xenopus oocyte studies revealed that AtAVT6D is involved in the uptake of Aspartic acid. While overexpression of AtAVT6D resulted in smaller siliques in Arabidopsis thaliana. Additionally, transient expression studies were performed with the full-length AtAVT6D promoter and its deletion constructs to study the effect of ACGT-N
Identifiants
pubmed: 35377091
doi: 10.1007/s11103-022-01256-x
pii: 10.1007/s11103-022-01256-x
doi:
Substances chimiques
Aspartic Acid
30KYC7MIAI
Abscisic Acid
72S9A8J5GW
Nitrogen
N762921K75
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
67-82Subventions
Organisme : Science and Engineering Research Board
ID : EMR/2016/002470
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/P012523/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/P012574/1
Pays : United Kingdom
Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature B.V.
Références
Amir R, Galili G, Cohen H (2018) The metabolic roles of free amino acids during seed development. Plant Sci 275:11–18. https://doi.org/10.1016/j.plantsci.2018.06.011
doi: 10.1016/j.plantsci.2018.06.011
pubmed: 30107877
Angelovici R, Fait A, Zhu X, Szymanski J, Feldmesser E, Fernie AR, Galili G (2009) Deciphering transcriptional and metabolic networks associated with lysine metabolism during Arabidopsis seed development. Plant Physiol 151:2058–2072. https://doi.org/10.1104/PP.109.145631
doi: 10.1104/PP.109.145631
pubmed: 19783646
pmcid: 2785976
Angelovici R, Fait A, Fernie AR, Galili G (2011) A seed high-lysine trait is negatively associated with the TCA cycle and slows down Arabidopsis seed germination. New Phytol 189:148–159. https://doi.org/10.1111/J.1469-8137.2010.03478.X
doi: 10.1111/J.1469-8137.2010.03478.X
pubmed: 20946418
Balazadeh S, Schildhauer J, Araújo WL, Munné-Bosch S, Fernie AR, Proost S, Humbeck K, Mueller-Roeber B (2014) Reversal of senescence by N resupply to N-starved Arabidopsis thaliana: transcriptomic and metabolomic consequences. J Exp Bot 65:3975–3992. https://doi.org/10.1093/jxb/eru119
doi: 10.1093/jxb/eru119
pubmed: 24692653
pmcid: 4106441
Besnard J, Sonawala U, Maharjan B, Collakova E, Finlayson SA, Pilot G, McDowell J, Okumoto S (2021) Increased expression of UMAMIT amino acid transporters results in activation of salicylic acid dependent stress response. Front Plant Sci 11:2214. https://doi.org/10.3389/FPLS.2020.606386/BIBTEX
doi: 10.3389/FPLS.2020.606386/BIBTEX
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
doi: 10.1016/0003-2697(76)90527-3
pubmed: 942051
Chahomchuen T, Sekito T, Hondo K, Nishimoto S, Sugahara T, Kakinuma Y (2008) Characterization of the vacuolar transporters for amino acid recycling in yeast autophagy. Interdiscip Stud Environ Chem Biol Responses Chem Pollut 1:251–261
Chahomchuen T, Hondo K, Ohsaki M, Sekito T, Kakinuma Y (2009) Evidence for Avt6 as a vacuolar exporter of acidic amino acids in Saccharomyces cerevisiae cells. J Gen Appl Microbiol 55:409–417. https://doi.org/10.2323/jgam.55.409
doi: 10.2323/jgam.55.409
pubmed: 20118605
Chen L, Bush DR (1997) LHT1, a lysine- and histidine-specific amino acid transporter in Arabidopsis. Plant Physiol 115:1127–1134. https://doi.org/10.1104/pp.115.3.1127
doi: 10.1104/pp.115.3.1127
pubmed: 9390441
pmcid: 158577
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743. https://doi.org/10.1046/J.1365-313X.1998.00343.X
doi: 10.1046/J.1365-313X.1998.00343.X
pubmed: 10069079
Dhatterwal P, Mehrotra S, Mehrotra R (2017) Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana. BMC Res Notes 10:638. https://doi.org/10.1186/s13104-017-2982-1
doi: 10.1186/s13104-017-2982-1
pubmed: 29183338
pmcid: 5706289
Dhatterwal P, Mehrotra S, Miller AJ, Mehrotra R (2021) Promoter profiling of Arabidopsis amino acid transporters: clues for improving crops. Plant Mol Biol 107:451–475. https://doi.org/10.1007/S11103-021-01193-1
doi: 10.1007/S11103-021-01193-1
pubmed: 34674117
Frankard V, Ghislain M, Jacobs M (1992) Two feedback-insensitive enzymes of the aspartate pathway in Nicotiana sylvestris. Plant Physiol 99:1285–1293. https://doi.org/10.1104/PP.99.4.1285
doi: 10.1104/PP.99.4.1285
pubmed: 16669034
pmcid: 1080622
Freeman J, Sparks CA, West J, Shewry PR, Jones HD (2011) Temporal and spatial control of transgene expression using a heat-inducible promoter in transgenic wheat. Plant Biotechnol J 9:788–796. https://doi.org/10.1111/J.1467-7652.2011.00588.X
doi: 10.1111/J.1467-7652.2011.00588.X
pubmed: 21265997
Frommer WB, Ninnemann O (1995) Heterologous expression of genes in bacterial, fungal, animal, and plant cells. Annu Rev Plant Physiol Plant Mol Biol 46:419–444. https://doi.org/10.1146/annurev.pp.46.060195.002223
doi: 10.1146/annurev.pp.46.060195.002223
Fujiki Y, Teshima H, Kashiwao S, Kawano-Kawada M, Ohsumi Y, Kakinuma Y, Sekito T (2017) Functional identification of AtAVT3, a family of vacuolar amino acid transporters, in Arabidopsis. FEBS Lett 591:5–15. https://doi.org/10.1002/1873-3468.12507
doi: 10.1002/1873-3468.12507
pubmed: 27925655
Galili G (2011) The aspartate-family pathway of plants: Linking production of essential amino acids with energy and stress regulation. Plant Signal Behav 6:192–195. https://doi.org/10.4161/psb.6.2.14425
doi: 10.4161/psb.6.2.14425
pubmed: 21512320
pmcid: 3121977
Galili G, Amir R (2013) Fortifying plants with the essential amino acids lysine and methionine to improve nutritional quality. Plant Biotechnol J 11:211–222. https://doi.org/10.1111/pbi.12025
doi: 10.1111/pbi.12025
pubmed: 23279001
Galili G, Avin-Wittenberg T, Angelovici R, Fernie AR (2014) The role of photosynthesis and amino acid metabolism in the energy status during seed development. Front Plant Sci 5:447. https://doi.org/10.3389/fpls.2014.00447
doi: 10.3389/fpls.2014.00447
pubmed: 25232362
pmcid: 4153028
Gietz RD, Woods RA (2002) Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. Methods Enzymol 350:87–96. https://doi.org/10.1016/S0076-6879(02)50957-5
doi: 10.1016/S0076-6879(02)50957-5
pubmed: 12073338
Grunennvaldt RL, Degenhardt-Goldbach J, Gerhardt IR, Quoirin M (2015) Promoters used in genetic transformation of plants. Res J Biol Sci 10:1–9. https://doi.org/10.3923/rjbsci.2015.1.9
doi: 10.3923/rjbsci.2015.1.9
Helm CV, De Francisco A, Gaziola SA, Fornazier RF, Pompeu GB, Azevedo RA (2004) Hull-less barley varieties: storage proteins and amino acid distribution in relation to nutritional quality. Food Biotechnol 18:327–341. https://doi.org/10.1081/FBT-200040531
doi: 10.1081/FBT-200040531
Hildebrandt TM, Nunes Nesi A, Araújo WL, Braun HP (2015) Amino acid catabolism in plants. Mol Plant 8:1563–1579. https://doi.org/10.1016/j.molp.2015.09.005
doi: 10.1016/j.molp.2015.09.005
pubmed: 26384576
Hirota T, Izumi M, Wada S, Makino A, Ishida H (2018) Vacuolar protein degradation via autophagy provides substrates to amino acid catabolic pathways as an adaptive response to sugar starvation in Arabidopsis thaliana. Plant Cell Physiol 59:1363–1376. https://doi.org/10.1093/pcp/pcy005
doi: 10.1093/pcp/pcy005
pubmed: 29390157
Huang A, Coutu C, Harrington M, Rozwadowski K, Hegedus DD (2021) Engineering a feedback inhibition-insensitive plant dihydrodipicolinate synthase to increase lysine content in Camelina sativa seeds. Transgenic Res. https://doi.org/10.1007/S11248-021-00291-6
doi: 10.1007/S11248-021-00291-6
pubmed: 34802109
pmcid: 8821502
Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Report 5:387–405. https://doi.org/10.1007/BF02667740
doi: 10.1007/BF02667740
Jia M, Wu H, Clay KL, Jung R, Larkins BA, Gibbon BC (2013) Identification and characterization of lysine-rich proteins and starch biosynthesis genes in the opaque2 mutant by transcriptional and proteomic analysis. BMC Plant Biol 13:60. https://doi.org/10.1186/1471-2229-13-60
doi: 10.1186/1471-2229-13-60
pubmed: 23586588
pmcid: 3762070
Kirma M, Araújo WL, Fernie AR, Galili G (2012) The multifaceted role of aspartate-family amino acids in plant metabolism. J Exp Bot 63:4995–5001. https://doi.org/10.1093/jxb/ers119
doi: 10.1093/jxb/ers119
pubmed: 22516796
Lee YH, Foster J, Chen J, Voll LM, Weber APM, Tegeder M (2007) AAP1 transports uncharged amino acids into roots of Arabidopsis. Plant J 50:305–319. https://doi.org/10.1111/J.1365-313X.2007.03045.X
doi: 10.1111/J.1365-313X.2007.03045.X
pubmed: 17419840
Liang G, He H, Yu D (2012) Identification of nitrogen starvation-responsive microRNAs in Arabidopsis thaliana. PLoS ONE 7:e48951. https://doi.org/10.1371/journal.pone.0048951
doi: 10.1371/journal.pone.0048951
pubmed: 23155433
pmcid: 3498362
Lim PO, Kim HJ, Gil Nam H (2007) Leaf senescence. Annu Rev Plant Biol 58:115–136. https://doi.org/10.1146/annurev.arplant.57.032905.105316
doi: 10.1146/annurev.arplant.57.032905.105316
pubmed: 17177638
Liu JH, Peng T, Dai W (2014) Critical cis-acting elements and interacting transcription factors: key players associated with abiotic stress responses in plants. Plant Mol Biol Report 32:303–317. https://doi.org/10.1007/s11105-013-0667-z
doi: 10.1007/s11105-013-0667-z
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
doi: 10.1006/meth.2001.1262
pubmed: 11846609
Locascio A, Andrés-Colás N, Mulet JM, Yenush L (2019) Saccharomyces cerevisiae as a tool to investigate plant potassium and sodium transporters. Int J Mol Sci 20:2133. https://doi.org/10.3390/ijms20092133
doi: 10.3390/ijms20092133
pmcid: 6539216
Lu K, Wu B, Wang J, Zhu W, Nie H, Qian J, Huang W, Fang Z (2018) Blocking amino acid transporter OsAAP3 improves grain yield by promoting outgrowth buds and increasing tiller number in rice. Plant Biotechnol J 16:1710–1722. https://doi.org/10.1111/pbi.12907
doi: 10.1111/pbi.12907
pubmed: 29479779
pmcid: 6131477
Mehrotra R, Mehrotra S (2010) Promoter activation by ACGT in response to salicylic and abscisic acids is differentially regulated by the spacing between two copies of the motif. J Plant Physiol 167:1214–1218. https://doi.org/10.1016/j.jplph.2010.04.005
doi: 10.1016/j.jplph.2010.04.005
pubmed: 20554077
Mehrotra R, Kiran K, Chaturvedi CP, Ansari SA, Lodhi N, Sawant S, Tuli R (2005) Effect of copy number and spacing of the ACGT and GT cis elements on transient expression of minimal promoter in plants. J Genet 84:183–187. https://doi.org/10.1007/BF02715844
doi: 10.1007/BF02715844
pubmed: 16131718
Mehrotra R, Yadav A, Bhalothia P, Karan R, Mehrotra S (2012) Evidence for directed evolution of larger size motif in Arabidopsis thaliana genome. Sci World J. https://doi.org/10.1100/2012/983528
doi: 10.1100/2012/983528
Mehrotra R, Sethi S, Zutshi I, Bhalothia P, Mehrotra S (2013) Patterns and evolution of ACGT repeat cis-element landscape across four plant genomes. BMC Genomics 14:203. https://doi.org/10.1186/1471-2164-14-203
doi: 10.1186/1471-2164-14-203
pubmed: 23530833
pmcid: 3622567
Miller AJ, Zhou JJ (2000) Xenopus oocytes as an expression system for plant transporters. Biochim Biophys Acta Biomembr 1465:343–358. https://doi.org/10.1016/S0005-2736(00)00148-6
doi: 10.1016/S0005-2736(00)00148-6
Potenza C, Aleman L, Sengupta-Gopalan C (2004) Targeting transgene expression in research, agricultural, and environmental applications: promoters used in plant transformation. Vitr Cell Dev Biol Plant 40:1–22. https://doi.org/10.1079/IVP2003477
doi: 10.1079/IVP2003477
Rentsch D, Hirner B, Schmelzer E, Frommer WB (1996) Salt stress-induced proline transporters and salt stress-repressed broad specificity amino acid permeases identified by suppression of a yeast amino acid permease-targeting mutant. Plant Cell 8:1437–1446. https://doi.org/10.1105/tpc.8.8.1437
doi: 10.1105/tpc.8.8.1437
pubmed: 8776904
pmcid: 161269
Rushton PJ, Reinstädler A, Lipka V, Lippok B, Somssich IE (2002) Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen-and wound-induced signaling. Plant Cell 14:749–762. https://doi.org/10.1105/tpc.010412
doi: 10.1105/tpc.010412
pubmed: 11971132
pmcid: 150679
Russnak R, Konczal D, McIntire SL (2001) A family of yeast proteins mediating bidirectional vacuolar amino acid transport. J Biol Chem 276:23849–23857. https://doi.org/10.1074/jbc.M008028200
doi: 10.1074/jbc.M008028200
pubmed: 11274162
Sah SK, Reddy KR, Li J (2016) Abscisic acid and abiotic stress tolerance in crop plants. Front Plant Sci 7:571. https://doi.org/10.3389/fpls.2016.00571
doi: 10.3389/fpls.2016.00571
pubmed: 27200044
pmcid: 4855980
Schroeder JI, Delhaize E, Frommer WB, Lou GM, Harrison MJ, Herrera-Estrella L, Horie T, Kochian LV, Munns R, Nishizawa NK, Tsay YF, Sanders D (2013) Using membrane transporters to improve crops for sustainable food production. Nature 497:60–66. https://doi.org/10.1038/nature11909
doi: 10.1038/nature11909
pubmed: 23636397
pmcid: 3954111
Seifi HS, Van BJ, Angenon G, Höfte M (2013) Glutamate metabolism in plant disease and defense: friend or foe? MPMI 26:475–485. https://doi.org/10.1094/MPMI-07-12-0176-CR
doi: 10.1094/MPMI-07-12-0176-CR
pubmed: 23342972
Seki M, Ishida J, Narusaka M, Fujita M, Nanjo T, Umezawa T, Kamiya A, Nakajima M, Enju A, Sakurai T, Satou M (2002) Monitoring the expression pattern of around 7,000 Arabidopsis genes under ABA treatments using a full-length cDNA microarray. Funct Integr Genom 2:282–291. https://doi.org/10.1007/S10142-002-0070-6
doi: 10.1007/S10142-002-0070-6
Shaul O, Galili G (1993) Concerted regulation of lysine and threonine synthesis in tobacco plants expressing bacterial feedback-insensitive aspartate kinase and dihydrodipicolinate synthase. Plant Mol Biol 23:759–768. https://doi.org/10.1007/BF00021531
doi: 10.1007/BF00021531
pubmed: 8251629
Shinozaki K, Yamaguchi-Shinozaki K (2003) Molecular mechanisms of plant responses and tolerance of drought and cold stress. Plant biotechnology 2002 and beyond. Springer, Dordrecht, pp 30–37
doi: 10.1007/978-94-017-2679-5_6
Wan J, Tanaka K, Zhang XC, Son GH, Brechenmacher L, Nguyen THN, Stacey G (2012) LYK4, a lysin motif receptor-like kinase, is important for chitin signaling and plant innate immunity in Arabidopsis. Plant Physiol 160:396–406. https://doi.org/10.1104/pp.112.201699
doi: 10.1104/pp.112.201699
pubmed: 22744984
pmcid: 3440214
Wang W, Vinocur B, Altman A (2003) Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14. https://doi.org/10.1007/s00425-003-1105-5
doi: 10.1007/s00425-003-1105-5
pubmed: 14513379
Wu S-J, Wang L-C, Yeh C-H, Lu C-A, Wu S-J (2010) Isolation and characterization of the Arabidopsis heat-intolerant 2 (hit2) mutant reveal the essential role of the nuclear export receptor EXPORTIN1A (XPO1A) in plant heat tolerance. New Phytol 186:833–842. https://doi.org/10.1111/J.1469-8137.2010.03225.X
doi: 10.1111/J.1469-8137.2010.03225.X
pubmed: 20345641
Xia T, Xiao D, Liu D, Chai W, Gong Q, Wang NN (2012) Heterologous expression of ATG8c from soybean confers tolerance to nitrogen deficiency and increases yield in Arabidopsis. PLoS ONE 7:e37217. https://doi.org/10.1371/journal.pone.0037217
doi: 10.1371/journal.pone.0037217
pubmed: 22629371
pmcid: 3358335
Yang H, Postel S, Kemmerling B, Ludewig U (2014) Altered growth and improved resistance of Arabidopsis against Pseudomonas syringae by overexpression of the basic amino acid transporter AtCAT1. Plant Cell Environ 37:1404–1414. https://doi.org/10.1111/pce.12244
doi: 10.1111/pce.12244
pubmed: 24895758
Yang QQ, Zhao DS, Zhang CQ, Wu HY, Li QF, Gu MH, Sun SSM, Liu QQ (2018) A connection between lysine and serotonin metabolism in rice endosperm. Plant Physiol 176:1965–1980. https://doi.org/10.1104/PP.17.01283
doi: 10.1104/PP.17.01283
pubmed: 29363563
pmcid: 5841688
Yang G, Wei Q, Huang H, Xia J (2020) Amino acid transporters in plant cells: a brief review. Plants 9:1–17. https://doi.org/10.3390/plants9080967
doi: 10.3390/plants9080967
Yu S, Pratelli R, Denbow C, Pilot G (2015) Suppressor mutations in the Glutamine Dumper1 protein dissociate disturbance in amino acid transport from other characteristics of the Gdu1D phenotype. Front Plant Sci 6:593. https://doi.org/10.3389/FPLS.2015.00593/BIBTEX
Zhang R, Zhu J, Cao HZ, Xie XL, Huang JJ, Chen XH, Luo ZY (2013) Isolation and characterization of LHT-type plant amino acid transporter gene from Panax ginseng Meyer. J Ginseng Res 37:361–370. https://doi.org/10.5142/jgr.2013.37.361
doi: 10.5142/jgr.2013.37.361
pubmed: 24198663
pmcid: 3818964