Abstract
Heat shock protein (HSP101) function as molecular chaperones and confer thermotolerance to plants. In the present investigation, identification, comprehensive expression analysis, phylogeny and protein modelling of HSP101 gene has been done in Aegilops speltoides accession Pau3583. In the present study, we cloned and in silico characterized a HSP101C gene designated as AsHSP101C-Pau3583. AsHSP101C-Pau3583 is 4180 bp long with seven exons and six introns and encoded a polypeptide of 910 amino acids predicted by FGENESH. We have identified 58 SNPs between the AsHSP101C-Pau3583 and reference gene sequence extracted from Ae. speltoides TGAC assembly. Real-time RT-PCR analysis of expression levels of HSP101 gene in two wheat genotypes under heat stress revealed that gene namely HSP101C was up-regulated in Aegilops speltoides acc. Pau3583 by > fourfold in comparison to Triticum aestivum cv. PBW343 under heat stress signifies that it plays a role in conferring heat tolerance. Sequence comparison and phylogenetic analysis of AsHSP101C-Pau3583 with seven wheat homologs Triticum aestivum, Aegilops speltoides (TGAC), Triticum durum cv Cappelli, Triticum durum cv Strongfield, Triticum monococcum, Aegilops tauschii and Triticum urartu showed significant similarities with highly conserved coding regions and functional domains (AAA, AAA + 2, ClpB domains), suggesting the conserved function of HSP101C in different species. The illustration of the protein models of HSP101C in homologs provided information for the ATP-binding motifs within the nucleotide binding domains (NBD), specific for the chaperone activity. These findings are important and identified SNPs could be used for designing markers for ensuring the transfer of AsHSP101C-Pau3583 gene into hexaploid wheat and its role in heat tolerance.
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Gene sequence submitted to NCBI with accession number KT867225.
Abbreviations
- HSP:
-
Heat shock protein
- NBD:
-
Nucleotide binding domains
- ClpB:
-
Caseinolytic protease domains
References
Agarwal M, Katiyar-Agarwal S, Sahi C, Gallie DR, Grover A (2001) Arabidopsis thaliana Hsp100 proteins: kith and kin. Cell Stress Chaperon 6:219–224
Agarwal M, Katiyar-Agarwal S, Grover A (2002) Plant Hsp100 proteins: structure, function and regulation. Plant Sci 163:397–405. https://doi.org/10.1016/S0168-9452(02)00209-1
Agarwal M, Sahi C, Katiyar Agarwal S, Agarwal S, Young T, Gallie TR, Sharma VM, Ganesan K, Grover A (2003) Molecular characterization of rice hsp101: complementation of yeast hsp104 mutation by disaggregation of protein granules and differential expression in indica and japonica rice type. Plant Mol Biol 51:543–553. https://doi.org/10.1023/A:1022324920316
Akter N, Islam MR (2017) Heat stress effects and management in wheat. A review. Agron Sustain Dev. https://doi.org/10.1007/s13593-017-0443-9
Almeselmani M, Deshmukh PS and Chinnusamy V (2011) Effects of prolonged high temperature stress on respiration, photosynthesis and gene expression in wheat (Triticum aestivum L.) varieties differing in their thermotolerance. Plant stress
Arora S, Cheema J, Poland J, Uauy C, Chhuneja P (2019) Genome-Wide association mapping of grain micronutrients concentration in Aegilops tauschii. Front Plant Sci 10:54. https://doi.org/10.3389/fpls.2019.00054
Asseng S, Ewert F, Martre P, Rötter RP, Lobell DB, Cammarano D (2015) Rising temperatures reduce global wheat production. Nat. Clim. Change 5:143–147. https://doi.org/10.1038/nclimate2470
Awlachew ZT, Singh R, Kaur S, Bains NS, Chhuneja P (2016) Transfer and mapping of the heat tolerance component traits of Aegilops speltoides in tetraploid wheat Triticum durum. Mol Breeding. https://doi.org/10.1007/s11032-016-0499-2
Barnett ME, Zolkiewska A, Zolkiewska M (2000) Structure and activity of ClpB from Escherichia coli. Role of the amino- and -carboxyl-terminal domains. J BioChem 275:37565–37571. https://doi.org/10.1074/jbc.M005211200
Campbell JL, Klueva NY, Zheng H, Nieto-Sotelo J, Ho THD, Nguyen HT (2001) Cloning of new members of heat shock protein HSP101 gene family in wheat (Triticum aestivum (L.) Moench) inducible by heat, dehydration, and ABA. Biochem Biophys Acta 1517:270–277. https://doi.org/10.1016/S0167-4781(00)00292-X
Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DJ, Thompson JD (2003) Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31:3497–3500. https://doi.org/10.1385/1-59259-890-0:493
Efeolgu B, Terzioglu S (2009) Photosynthetic responses of two wheat varieties to high temperature. EurAsia J BioSci 3:97–106. https://doi.org/10.5053/ejobios.2009.3.0.13
Erdayani E, Nagarajan R, Grant NP, Gill KS (2020) Genome-wide analysis of the HSP101/CLPB gene family for heat tolerance in hexaploid wheat. Sci Rep 10:3948. https://doi.org/10.1038/s41598-020-60673-4
Finn RD, Bateman A, Clements J, Coggill P, Eberhardt RY, Eddy SR, Heger A, Hetherington K, Holm L, Mistry J, Sonnhammer ELL, Tate J, Punta M (2014) Pfam: the protein families database. Nucleic Acids Res. https://doi.org/10.1093/nar/gkt1223
Gottesman S, Squires C, Pichersky E, Carrington M, Hobbs M, Mattick JS, Dalrymple B, Kuramitsu H, Shiroza T, Foster T (1990) Conservation of the regulatory subunit for the Clp ATP-dependent protease in prokaryotes and eukaryotes. Proc Natl AcadSci USA 87:3513–3517. https://doi.org/10.1073/pnas.87.9.3513
Gulli M, Corradi M, Rampino P, Marmiroli N, Perrotta C (2007) Four members of the HSP101 gene family are differently regulated in Triticum durum Desf. FEBS Lett 581:4841. https://doi.org/10.1016/j.febslet.2007.09.010
Hairat S, Khurana P (2016) Photosynthetic efficiency, temperature induction response, carbon isotope discrimination correlate with expression profiling in Indian wheat cultivars. Plant Signal Behav 11:9. https://doi.org/10.1080/15592324.2016.1179416
He Y, Liu X, Huang B (2005) Protein changes in response to heat stress in acclimated and nonacclimated creeping bentgrass. J Amer Soc Hort Sci 130:521–526. https://doi.org/10.21273/JASHS.130.4.521
Hendershot KL Weng J and Nguyen HT (1992) Induction Temperature of Heat‐Shock Protein Synthesis in Wheat. Crop Sci. 32(1):256-261. https://doi.org/10.2135/cropsci1992.0011183X003200010051x
Hong SW, Vierling E (2001) HSP101 is necessary for heat tolerance but dispensable for development and germination in the absence of stress. Plant J 27:25–35. https://doi.org/10.1046/j.1365-313x.2001.01066.x
Katiyar-Agarwal S, Agarwal M, Grover A (2003) Heat tolerant basmati rice engineered by over-expression of hsp 101. Plant Mol Biol 51:677–686
Koonin EV, Galperin MY. (2003) Sequence - Evolution – Function-Computational approaches in Comparative genomics Evolutionary Concept in Genetics and Genomics. 2003 edition, Springer, 1–40207–274–0
Kumar S, Kumara P, Kumar U, Grover M, Singh AK, Singh R, Senger RS (2013) Molecular approaches for designing heat tolerant wheat. J Plant Biochem Biotechnol 22:359–371. https://doi.org/10.1007/s13562-013-0229-3
Larkindale J, Hall JD, Knight MR, Vierling E (2005) Heat stress phenotypes of Arabidopsis mutants implicate multiple signaling pathways in the acquisition of thermotolerance. Plant Physiol 138:882–897. https://doi.org/10.1104/pp.105.062257
Lee U, Rioflorido I, Hong SW, Larkindale J, Waters ER, Vierling E (2007) The Arabidopsis ClpB/Hsp100 family of proteins: chaperones for stress and chloroplast development. Plant J 49:115–127. https://doi.org/10.1111/j.1365-313X.2006.02940.x
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-∆∆CT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Maroof S (1984) Ribosomal DNA spacer length polymorphism in barley: Mendelian inheritance, chromosomal location and population dynamics. Proc Natl Acad Sci USA 81:8014–8018
Muthusamy SK, Dalal M, Chinnusamy V, Bansal KC (2016) Differential regulation of genes coding for organelle and cytosolic ClpATPases under biotic and abiotic stresses in wheat. Front Plant Sci 7:929. https://doi.org/10.3389/fpls.2016.00929
Nayak SN, Balaji J, Upadhyaya HD, Hash CT, Kishor PBK, Chattopadhyay D (2009) Isolation and sequence analysis of DREB2A homologues in three cereal and two legume species. Plant Sci 177:460–467. https://doi.org/10.1016/JPlantSci.2009.07.009
Nieto-Sotelo J, Martínez LM, Ponce G, Cassab GI, Alagón A, Meeley RB (2002) Maize HSP101 plays important roles in both induced and basal thermotolerance and primary root growth. Plant Cell 14:1621–1633. https://doi.org/10.1105/tpc.010487
Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE (2004) UCSF Chimera–a visualization system for exploratory research and analysis. J Comput Chem 25:1605–1612. https://doi.org/10.1002/jcc.20084
Pradhan GP, Prasad PVV, Fritz AK, Kirkham MB, Gill BS (2012) High temperature tolerance in Aegilops species and its potential transfer to wheat. Crop Sci 52:292–304. https://doi.org/10.2135/cropsci2011.04.0186
Prajapati C, Bhagat C (2012) In-silico analysis and homology modeling of target proteins for Clostridium Botulinum. Int J Pharm Sci Res 3:2050–2056
Rampino P, Mita G, Fasano P, Borrelli GM, Aprile A, Dalessandro G (2012) Novel durum wheat genes up-regulated in response to a combination of heat and drought stress. Plant Physiol Biochem 56:72–78. https://doi.org/10.1016/j.plaphy.2012.04.006
Ray DK, Mueller ND, West PC, Foley JA (2013) Yield trends are insufficient to double global crop production by 2050. PLoS ONE. https://doi.org/10.1371/journal.pone.0066428
Rout SB, Mishra S (2012) Protein structure prediction using stochastic process probabilistic model. Int J of Innovative Res in Sci, Eng and Tech 1:2
Rutherford K, Parkhill J, Crook J, Horsnell T, Rice P, Rajandrem MA, Bart B (2000) Artemis:sequence visualization and annotation. Bioinfo 16:944–945. https://doi.org/10.1093/bioinformatics/16.10.944
Sauer RT, Bolon DN, Burton BM, Burton RE, Flynn JM, Grant RA, Hersch GL, Joshi SA, Kenniston JA, Levchenko I, Neher SB, Oakes ESC, Siddiqui SM, Wah DA, Baker TA (2004) Sculpting the proteome with AAA+ proteases and dissembly machines. Cell 119:9–18. https://doi.org/10.1016/j.cell.2004.09.020
Schirmer EC, Lindquist S, Vierling E (1994) An Arabidopsis heat shock protein complements a thermotolerance defect in yeast. Plant Cell 6:1899–1909
Schirmer EC, Glover JR, Singer MA, Lindquist S (1996) Hsp100/Clp proteins: a common mechanism explains diverse functions. Trends BiochemSci 21:289–296. https://doi.org/10.1016/S0968-0004(96)10038-4
Singh A, Grover A (2010) Plant Hsp100/ClpB-like proteins: poorly analyzed cousins of yeast ClpB machine. Plant Mol Biol 74:395–404. https://doi.org/10.1007/s11103-010-9682-8
Singh SP, Sharma V (2017) In silico studies of conformational changes in wheat heat shock proteins. MOJ Proteom and Bioinform. 5(1):26–29
Singh A, Singh U, Mittal D, Grover A (2010) Genome-wide analysis of rice ClpB/HSP100 ClpC and ClpD genes. BMC Genom 11:95. https://doi.org/10.1186/1471-216411-95
Skylas DJ, Cordwell SJ, Hains PG, Larsen MR, Basseal DJ, Walsh BJ, Blumenthal C, Rathmell W, Copeland L, Wrigley CW (2002) Heat shock of Wheat during grain filling: proteins associated with heat-tolerance. J Cereal Sci 35:175–188. https://doi.org/10.1006/jcrs.2001.0410
Stone P (2001) The effects of heat stress on cereal yield and quality. In: Basra AS (ed) Crop responses and adaptation to temperature stress. Food Products Press, Binghamton, NY, pp 243–291
Trosch R, Mühlhaus T, Schroda M, Willmund F (2015) ATP-dependent molecular chaperones in plastids more complex than expected. Biochim Biophys Acta 1847:872–888. https://doi.org/10.1016/j.bbabio.2015.01.002
Webb B, Sali A (2014) Protein structure modeling with MODELLER. Methods Mol Bio 11:1–15. https://doi.org/10.1007/978-1-4939-0366-5_1
Yang J, Sun Y, Sun A, Yi S, Qin J, Li M (2006) The involvement of chloroplast HSP100/ClpB in the acquired thermotolerance in tomato. Plant Mol Biol 62:385–395. https://doi.org/10.1007/s11103-006-9027-9
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(information that explains whether and by whom the research was supported) The authors gratefully acknowledge the financial support by Department of Biotechnology, Government of India under the grant BT/PR7851/AGR/2/376/2006.
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Conceptualization and methodology by KS. Material preparation, data collection, wet lab experiments and data analysis were performed by PJ. KS and PC conceived the study, provided genetic material, arranged funding and helped in manuscript writing. SK provided technical assistance: PJ, IY did the bioinformatics work. PJ wrote the manuscript; PJ, PS, IY, PK edit the manuscript; KS and PC review and did final editing. All authors commented on previous versions of the manuscript. All authors have critically read and approved final manuscript.
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Jakhu, P., Sharma, P., Yadav, I.S. et al. Cloning, expression analysis and In silico characterization of HSP101: a potential player conferring heat stress in Aegilops speltoides (Tausch) Gren. Physiol Mol Biol Plants 27, 1205–1218 (2021). https://doi.org/10.1007/s12298-021-01005-2
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DOI: https://doi.org/10.1007/s12298-021-01005-2