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Abiotic stress regulates expression of galactinol synthase genes post-transcriptionally through intron retention in rice

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Abstract

Main conclusion

Expression of the Galactinol synthase genes in rice is regulated through post-transcriptional intron retention in response to abiotic stress and may be linked to Raffinose Family Oligosaccharide synthesis in osmotic perturbation.

Galactinol synthase (GolS) is the first committed enzyme in raffinose family oligosaccharide (RFO) synthesis pathway and synthesizes galactinol from UDP-galactose and inositol. Expression of GolS genes has long been implicated in abiotic stress, especially drought and salinity. A non-canonical regulation mechanism controlling the splicing and maturation of rice GolS genes was identified in rice photosynthetic tissue. We found that the two isoforms of Oryza sativa GolS (OsGolS) gene, located in chromosomes 3(OsGolS1) and 7(OsGolS2) are interspersed by conserved introns harboring characteristic premature termination codons (PTC). During abiotic stress, the premature and mature transcripts of both isoforms were found to accumulate in a rhythmic manner for very small time-windows interrupted by phases of complete absence. Reporter gene assay using GolS promoters under abiotic stress does not reflect this accumulation profile, suggesting that this regulation occurs post-transcriptionally. We suggest that this may be due to a surveillance mechanism triggering the degradation of the premature transcript preventing its accumulation in the cell. The suggested mechanism fits the paradigm of PTC-induced Nonsense-Mediated Decay (NMD). In support of our hypothesis, when we pharmacologically blocked NMD, the full-length pre-mRNAs were increasingly accumulated in cell. To this end, our work suggests that a combined transcriptional and post transcriptional control exists in rice to regulate GolS expression under stress. Concurrent detection and processing of prematurely terminating transcripts coupled to repressed splicing can be described as a form of Regulated Unproductive Splicing and Translation (RUST) and may be linked to the stress adaptation of the plant, which is an interesting future research possibility.

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References

  • Andersen KE, Bjergegaard C, Møller P, Sørensen JC, Sørensen H (2005) Compositional variations for alpha-galactosides in different species of leguminosae, brassicaceae, and barley: a chemotaxonomic study based on chemometrics and high-performance capillary electrophoresis. J Agric Food Chem 53:5809–5817

    Article  CAS  PubMed  Google Scholar 

  • Avigad G, Dey PM (1997) Carbohydrate metabolism: Storage carbohydrates. In: Dey PM, Harbourne J (eds) Plant biochemistry. Academic Press, San Diego, pp 143–204

    Chapter  Google Scholar 

  • Bachmann M, Matile P, Keller F (1994) Metabolism of the raffinose family oligosaccharides in leaves of Ajuga reptans L. Cold acclimation, translocation, sink to source transition: discovery of chain elongation enzyme. Plant Physiol 105:1335–1345

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Bringhurst RM, Cardon ZG, Gage DJ (2001) Galactosides in the rhizosphere: Utilization by Sinorhizobium meliloti and development of a biosensor. Proc Natl Acad Sci USA 98:4540–4545

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brown JWS (1986) A catalogue of splice junction and putative branchpoint sequences from plant introns. Nucleic Acids Res 14:9549–9559

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chang YF, Imam S, Wilkinsonv MF (2007) The nonsense-mediated decay RNA surveillance pathway. Annu Rev Biochem 76:51–74

    Article  CAS  PubMed  Google Scholar 

  • Cunningham SM, Nadeau P, Castonguay Y, Laberge S, Volenec JJ (2003) Raffinose and stachyose accumulation, galactinol synthase expression, and winter injury of contrasting alfalfa germplasms. Crop Sci 43:562–570

    Article  CAS  Google Scholar 

  • Demel RA, Dorrepaal E, Ebskamp MJM, Smeekens JCM, de Kruijff B (1998) Fructans interact strongly with model membranes. Biochim Biophys Acta 1375:36–42

    Article  CAS  PubMed  Google Scholar 

  • Dey PM (1985) D-Galactose-containing oligosaccharides. In: Dey PM, Dixon R (eds) Biochemistry of storage carbohydrates in green plants. Academic Press, San Diego, pp 53–129

    Google Scholar 

  • Dickey LF, Nguyen TT, Allen GC, Thompson WF (1994) Light modulation of ferredoxin mRNA abundance requires an open reading frame. Plant Cell 6:1171–1176

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • dos Santos TB, Budzinski IGF, Marur CJ, Petkowicz CLO, Pereira LFP, Vieira LGE (2011) Expression of three galactinol synthase isoforms in Coffea Arabica L. and accumulation of raffinose and stachyose in response to abiotic stresses. Plant Physiol Biochem 49:441–448

    Article  CAS  PubMed  Google Scholar 

  • dos Santos TB, de LimaRB Nagashima GT, de Oliveira Petkowicz CL, Carpentieri-Pípolo V, Protasio Pereira LF et al (2015) Galactinol synthase transcriptional profile in two genotypes of Coffea canephora with contrasting tolerance to drought. Genet Mol Biol 38(2):182–190

    Article  PubMed  PubMed Central  Google Scholar 

  • Downie B, Gurusinghe S, Dahal P, Thacker RR, Snyder JC, Nonogaki H et al (2003) Expression of a GALACTINOL SYNTHASE gene in tomato seeds is up-regulated before maturation desiccation and again after imbibition whenever radicle protrusion is prevented. Plant Physiol 131:1347–1359

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fan Y, Yu M, Liu M, Zhang R, Sun W, Qian M, et al (2017) Genome-wide identification, evolutionary and expression analyses of the GALACTINOL SYNTHASE gene family in rapeseed and tobacco. Int J Mol Sci 18(12):2768

    Article  CAS  PubMed Central  Google Scholar 

  • Filichkin SA, Mockler TC (2012) Unproductive alternative splicing and nonsense mRNAs: a widespread phenomenon among plant circadian clock genes. Biol Direct 7:20

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorus. J Biol Chem 66:375–400

    CAS  Google Scholar 

  • Gadjieva R, Axelsson E, Olsson U, Vallon-Christersson J, Hansson M (2004) Nonsense-mediated mRNA decay in barley mutants allows the cloning of mutated genes by a microarray approach. Plant Physiol Biochem 42:681–685

    Article  CAS  PubMed  Google Scholar 

  • Gallagher SR (1992) Quantitation of GUS activity by fluorometry. In: Gallagher SR (ed) GUS protocols: using the GUS gene as a reporter of gene expression. Academic Press, San Diego, pp 47–59

    Chapter  Google Scholar 

  • Gil P, Green PJ (1996) Multiple regions of the Arabidopsis SAUR-AC1 gene control transcript abundance: the 3′ untranslated region functions as an mRNA instability determinant. EMBO J 15(7):1678–1686

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gojłoa E, Pupel P, Lahuta LB, Podlinski P, Kucewicz M, Górecki RJ (2015) The acquisition of desiccation tolerance in developing Vicia hirsuta seeds coincides with an increase in galactinol synthase expression and soluble-d-galactosides accumulation. J Plant Physiol 184:37–48

    Article  CAS  Google Scholar 

  • Haab C, Keller F (2002) Purification and characterization of the raffinose oligosaccharide chain elongation enzyme, galactan: galactan galactosyltransferase (GGT), from Ajuga reptans leaves. Plant Physiol 114:361–371

    Article  CAS  Google Scholar 

  • Hannah MA, Zuther E, Buchel K, Heyer AG (2006) Transport and metabolism of raffinose family oligosaccharides in transgenic potato. J ExpBot 57(14):3801–3811

    CAS  Google Scholar 

  • Hao JH, Yang R, Fang KF, Wang JL, Zhang Q, Shen YY, Li TL (2014) Low night temperatures inhibit galactinol synthase gene expression and phloem loading in melon leaves during fruit development. Russ J Plant Physiol 61(2):178–187

    Article  CAS  Google Scholar 

  • Haritatos E, Medville R, Turgeon R (2000) Minor vein structure and sugar transport in Arabidopsis thaliana. Planta 211:105–111

    Article  CAS  PubMed  Google Scholar 

  • Isshiki M, Yamamoto Y, Satoh H, Shimamoto K (2001) Nonsense-mediated decay of mutant waxy mRNA in Rice. Plant Physiol 125:1388–1395

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jofuku KD, Schipper RD, Goldberg RB (1989) A frameshift mutation prevents Kunitz trypsin inhibitor mRNA accumulation in Soybean Embryos. Plant Cell 1:427–435

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kerenyi Z, Merai Z, Hiripi L, Benkovics A, Gyula P, Lacomme C et al (2008) Interkingdom conservation of mechanism of nonsense-mediated mRNA decay. EMBO J 27:1585–1595

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685

    Article  CAS  PubMed  Google Scholar 

  • Lareau LF, Brenner SE (2015) Regulation of splicing factors by alternative splicing and NMD is conserved between kingdoms yet evolutionarily flexible. Mol Biol Evol 32(4):1072–1079

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lemoine R, La Camera S, Atanassova R, Dédaldéchamp F, Allario T, Pourtau N et al (2013) Source-to-sink transport of sugar and regulation by environmental factors. Front Plant Sci 4:272

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li X, Zhuo J, Jing Y, Liu X, Wang X (2011) Expression of a Galactinol Synthase gene is positively associated with desiccation tolerance of Brassica napus seeds during development. J Plant Physiol 168:1761–1770

    Article  CAS  PubMed  Google Scholar 

  • Majee M, Maitra S, Dastidar KG, Pattnaik S, Chatterjee A, Hait NC, Das KP, Majumder AL (2004) A novel salt-tolerant l-myoinositol-1-phosphate synthase from Porteresia coarctata (Roxb.) Tateoka, a halophytic wild rice: molecular cloning, bacterial overexpression, characterization, and functional introgression into tobacco-conferring salt tolerance phenotype. J Biol Chem 279:28539–28552

    Article  CAS  PubMed  Google Scholar 

  • Manavella PA, Chan RL (2009) Transient transformation of sunflower leaf discs via an Agrobacterium-mediated method: applications for gene expression and silencing studies. Nat Prot 4(11):1699–1706

    Article  CAS  Google Scholar 

  • Marchant A, Bennett MJ (1998) The Arabidopsis AUX1 gene: a model system to study mRNA processing in plants. Plant Mol Biol 36:463–471

    Article  CAS  PubMed  Google Scholar 

  • McCaskill A, Turgeon R (2007) Phloem loading in Verbascum phoenicum L. depends on the synthesis of raffinose-family oligosaccharides. Proc Natl Acad Sci USA 104:1961–1962

    Article  Google Scholar 

  • Meyer T, Vigouroux A, Aumont-Nicaise M, Comte G, Vial L, Lavire C, Moréra S (2018) The plant defense signal galactinol is specifically used as a nutrient by the bacterial pathogen Agrobacterium fabrum. JBC Papers in Press. Published on March 30, 2018 as Manuscript RA118.001856

  • Miller MK, Schonhorst MH, McDaniel RG (1972) Identification of hybrids from alfalfa crosses by electrophoresis of single seed proteins1. Crop Science 12(4):535

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nishiguchi R, Takanami M, Oka A (1987) Characterization and sequence determination of the replicator region in the hairy-root inducing plasmid pRiA4b. Mol Gen Genet 206:1–8

    Article  CAS  Google Scholar 

  • Nishizawa A, Yabuta Y, Shigeoka S (2008) Galactinol and raffinose constitute a novel function to protect plants from oxidative damage. Plant Physiol 147:1251–1263

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Obendorf RL, Odorcic S, Ueda T, Coseo MP, Vassallo E (2004) Soybean galactinol synthase forms fagopyritol B1 but not galactopinitols: substrate feeding of isolated embryos and heterologous expression. Seed Sci Res 14:321–333

    Article  CAS  Google Scholar 

  • Panikulangara TJ, Eggers-Schumacher G, Wunderlich M, Stransky H, Schoffl F (2004) Galactinol synthase1: a novel heat shock factor target gene responsible for heat-induced synthesis of raffinose family oligosaccharides in Arabidopsis. Plant Physiol 136:3148–3158

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Passardi F, Penel C, Dunand C (2004) Performing the paradoxical: how plant peroxidases modify the cell wall. Trends Plant Sci 9:534–540

    Article  CAS  PubMed  Google Scholar 

  • Peshev D, Vergauwen R, Moglia A, Hideg E, Van den Ende W (2013) Towards understanding vacuolar antioxidant mechanisms: a role for fructans? J Exp Bot 64:1025–1038

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Peterbauer T, Richter A (2001) Biochemistry and physiology of raffinose family oligosaccharides and galactosyl cyclitols in seeds. Seed Sci Res 1:185–197

    Google Scholar 

  • Peters S, Mundree SG, Thomson JA, Farrant JM, Keller F (2007) Protection mechanisms in the resurrection plant Xerophyta viscosa (Baker): both sucrose and raffinose family oligosaccharides (RFOs) accumulate in leaves in response to water deficit. J Exp Bot 58:1947–1956

    Article  CAS  PubMed  Google Scholar 

  • Petracek ME, Nuygen T, Thompson WF, Dickey LF (2000) Premature termination codons destabilize ferredoxin-1 mRNA when ferredoxin-1 is translated. Plant J 21(6):563–569

    Article  CAS  PubMed  Google Scholar 

  • Que Q, Wang HY, English JJ, Jorgensen RA (1997) The frequency and degree of cosuppression by sense chalcone synthase transgenes are dependent on transgene promoter strength and are reduced by premature nonsense codons in the transgene coding sequence. Plant Cell 9:1357–1368

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rabbani MA, Maruyama K, Abe H, Khan MA, Katsura K, Ito Y et al (2003) Monitoring expression profiles of rice genes under cold, drought, and high-salinity stresses and abscisic acid application using cDNA microarray and RNA gel-blot Analyses. Plant Physiol 133:1755–1767

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ribeiro M, Felix CR, Lozzi SDP (2000) Soybean seed GALACTINOL SYNTHASE activity as determined by a novel colorimetric assay. R Bras Fisiol Veg 12(3):203–212

    Article  CAS  Google Scholar 

  • Ruijie F, Lixia H, Yang G, Qian M, Xin L (2017) Gene cloning and expression analysis of Galactinol Synthase gene VvGolS-4 in grape. Acta Agric Boreali-Sin 32(2):87–95

    Google Scholar 

  • Sahoo KK, Tripathi AK, Pareek A, Sopory SK, Singla-Pareek SL (2011) An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars. Plant Methods 7(1):49

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Saito M, Nakamura T (2005) Two point mutations identified in emmer wheat generate null Wx-A1 alleles. Theor Appl Genet 110:276–282

    Article  CAS  PubMed  Google Scholar 

  • Saito M, Yoshida M (2011) Expression analysis of the gene family associated with raffinose accumulation in rice seedlings under cold stress. Plant Physiol 168:2268–2271

    Article  CAS  Google Scholar 

  • Salvi P, Saxena SC, Petla BP, Kamble NU, Kaur H, Verma P et al (2016) Differentially expressed galactinol synthase(s) in chickpea are implicated in seed vigor and longevity by limiting the age induced ROS accumulation. Sci Rep 6:35088

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. In: Nolan C (ed) Laboratory manuals, 2nd edn. Cold Spring Harbor Laboratory Press, pp 7.39–7.52

  • Saravitz DM, Pharr DM, Carter TE (1987) Galactinol synthase activity and soluble sugars in developing seeds of four soybean genotypes. Plant Physiol 83:185–189

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sengupta S, Mukherjee S, Parween S, Majumder AL (2012) Galactinol synthase across evolutionary diverse taxa: functional preference for higher plants? FEBS Lett 586:1488–1496

    Article  CAS  PubMed  Google Scholar 

  • Shimosaka E, Ozawa K (2015) Overexpression of cold-inducible wheat galactinol synthase confers tolerance to chilling stress in transgenic rice. Breed Sci 65(5):363–371

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sikorskaite S, Rajamäki ML, Baniulis D, Stanys V, Valkonen JPT (2013) Protocol: optimised methodology for isolation of nuclei from leaves of species in the Solanaceae and Rosaceae families. Plant Methods 9:31

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Song J, Liu J, Weng M, Huang Y, Luo L, Cao P, Sun H et al (2013) Cloning of galactinol synthase gene from Ammopiptanthus mongolicus and its expression in transgenic Photinia serrulata plants. Gene 513:118–127

    Article  CAS  PubMed  Google Scholar 

  • Sprenger N, Keller F (2000) Allocation of raffinose family oligosaccharides to transport and storage pools in Ajuga reptans: the role of two distinct galactinol synthases. Plant J 21:249–258

    Article  CAS  PubMed  Google Scholar 

  • Stoyanova S, Geuns J, Hideg E, Van den Ende W (2011) The food additives inulin and stevioside counteract oxidative stress. Int J Food Sci Nutr 62:207–214

    Article  CAS  PubMed  Google Scholar 

  • Taji T, Ohsumi C, Iuchi S, Seki M, Kasuga M, Kobayashi M, Yamaguchi-Shinozaki K, Shinozaki K (2002) Important roles of drought- and cold-inducible genes for galactinol synthase in stress tolerance in Arabidopsis thaliana. Plant J 29(4):417–426

    Article  CAS  PubMed  Google Scholar 

  • Takanashi K, Shitan N, Sugiyama A, Kamimoto Y, Hamamoto M, Iwaki T et al (2008) Galactinol synthase gene of Coptis japonica is involved in Berberine Tolerance. Biosci Biotechnol Biochem 72(2):398–405

    Article  CAS  PubMed  Google Scholar 

  • Tapernoux-Lüthi EM, Schneider T, Keller F (2007) The C-terminal sequence from common bugle leaf galactan: galactan galactosyltransferase is an on-sequence-specific vacuolar sorting determinant. FEBS Lett 581:1811–1818

    Article  CAS  PubMed  Google Scholar 

  • Unda F, Canam T, Preston L, Mansfield SD (2011) Isolation and characterization of galactinol synthases from hybrid poplar. J Exp Bot 63(5):2059–2069

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Valluru R, Van den Ende W (2008) Plant fructans in stress environments: emerging concepts and future prospects. J Exp Bot 59:2905–2916

    Article  CAS  PubMed  Google Scholar 

  • Valluru R, Lammens W, Claupein W, Van den Ende W (2008) Freezing tolerance by vesicle-mediated fructan transport. Trends Plant Sci 13:409–414

    Article  CAS  PubMed  Google Scholar 

  • Van den Ende W, Valluru R (2009) Sucrose, sucrosyl oligosaccharides, and oxidative stress: scavenging and salvaging? J Exp Bot 60:9–18

    Article  CAS  PubMed  Google Scholar 

  • Vereyken IJ, ChupinV Demel RA, Smeekens SCM, DeKruijff B (2001) Fructans insert between the head groups of phospholipids. Biochim Biophys Acta Biomem 1510:307–320

    Article  CAS  Google Scholar 

  • Voelker TA, Moreno J, Chrispeels MJ (1990) Expression analysis of a pseudogene in transgenic tobacco: a frameshift mutation prevents mRNA accumulation. Plant Cell 2:255–261

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang Z, Zhu Y, Wang L, Liu X, Liu Y, Phillips J, Deng XA (2009) WRKY transcription factor participates in dehydration tolerance in Boea hygrometrica by binding to the W-box elements of the galactinol synthase (BhGolS1) promoter. Planta 230:1155–1166

    Article  CAS  PubMed  Google Scholar 

  • Zhao TY, Thacker R, Corum JW III, Synder JC, Meeley RB, Obendorf RL, Downie B (2004) Expression of the maize GALACTINOL SYNTHASE gene family: (I) Expression of two different genes during seed development and germination. Physiol Plantarum 121:634–646

    Article  CAS  Google Scholar 

  • Zhou T, Zhang R, Guo S (2012) Molecular cloning and characterization of GhGolS1, a novel gene encoding galactinol synthase from cotton (Gossypium hirsutum). Plant Mol Biol Rep 30:699–709

    Article  CAS  Google Scholar 

  • Zhou Y, Liu Y, Wang S, Shi C, Zhang R et al (2017) Molecular cloning and characterization of galactinol synthases in Camellia sinensis with different responses to biotic and abiotic stressors. J Agric Food Chem 65:2751–2759

    Article  CAS  PubMed  Google Scholar 

  • Zhuo C, Wanga T, Lua S, Zhaoa Y, Lia X, Guoa Z (2013) A cold responsive galactinol synthase gene from Medicago falcata (MfGolS1) is induced by myo-inositol and confers multiple tolerances to abiotic stresses. Physiol Plant 149:67–78

    Article  CAS  PubMed  Google Scholar 

  • Zuther E, Buchel K, Hundertmark M, Stitt M, Hincha DK, Heyer AG (2004) The role of raffinose in the cold acclimation response of Arabidopsis thaliana. FEBS Lett 576:169–173

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The project is supported by funds from Department of Biotechnology and the Raja Ramanna Fellowship Project of the Department of Atomic Energy, Government of India awarded to ALM, currently an INSA Senior Scientist. SM was supported by Fellowship from the Council of Scientific and Industrial Research (CSIR) and Department of Atomic Energy (DAE). SS was a Staff Scientist supported by DBT and Fast-Track young scientist scheme of Department of Science and Technology (DST), Government of India. AM is supported by the DBT project.

Funding

This study is funded by grants from the Department of Biotechnology (DBT654BT/AB/05/02/2007-III) and the Raja Ramanna Fellowship Project of the Department of Atomic Energy (D.O No. 10/25/2010/RRF-R & D-II/3118), both from the Government of India, awarded to ALM, currently an INSA Senior Scientist. SM and PB was supported by Fellowship from the Council of Scientific and Industrial Research (CSIR) and Department of Atomic Energy (DAE). AM is supported by the DBT project. SSG was a Staff Scientist supported by DBT and Fast-Track Young Scientist Scheme of Department of Science and Technology (DST), Government of India.

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  1. Sritama Mukherjee

    Present address: Botany Department, Bethune College, Kolkata, West Bengal, 700006, India

  2. Sonali Sengupta

    Present address: School of Plant Environment and Soil Sciences, LSUAg Center, Baton Rouge, LA, 70803, USA

  3. Sritama Mukherjee and Sonali Sengupta contributed equally.

Authors and Affiliations

  1. Division of Plant Biology, Bose Institute (Centenary Campus), Kolkata, West Bengal, 700054, India

    Sritama Mukherjee, Sonali Sengupta, Abhishek Mukherjee, Papri Basak & Arun Lahiri Majumder

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Electronic supplementary material

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Online Resource 1

(ESM_1.pdf): Primers and their sequences (PDF 89 kb)

Online Resource 2

(ESM_2.pdf): Splice site analysis of GolS genes in rice and wild rice relatives. Os: Oryza sativa, Ob: Oryza brachyantha, Om: Oryza minuta (PDF 90 kb)

Online Resource 3

(ESM_3.pdf): Sequence alignment of OsGolS with wild rice relatives. Exon-exon junctions are represented with black boxes. Os: Oryza sativa, Ob: Oryza brachyantha, Or: Oryza rufipogon, Om: Oryza minuta (TIFF 2668 kb)

Online Resource 4

(ESM_4.tif): a PCR amplification of GolS genes from cDNA of Porteresia coarctata (PcGolS1/PcGolS2) and Oryza sativa (OsGolS1/OsGolS2). Sequence alignment of OsGolS1 and PcGolS1 (b) and OsGolS2 and PcGolS2 (c) showing complete identity (TIFF 1585 kb)

Online Resource 5

(ESM_5.tif): Semiquantitative RT-PCR of (a) OsGolS1 and (b) OsGolS2 in one month old seedlings. Red triangles show occurrence of pre-mRNA, green triangles show occurrence of mature mRNA. Days indicate the stress treatment. c Quantitative PCR for all the above conditions. ‘*’ represents p-value < 0.05 as calculated through student’s t-test. (TIFF 1143 kb)

Online Resource 6

(ESM_6.tif): a–b Searching for an intronless OsGolS from different tissues. Pre-mRNAs were amplified only in mature leaves and germinated seedlings. c Quantitative PCR for all the above conditions. ‘*’ represents p value < 0.05 as calculated through student’s t-test (TIFF 610 kb)

Online Resource 7

(ESM_7.tif): In planta expression of OsGolS proteins. a Schematic diagram of binary vector pCAMBIA-1301 construct used in transformation. Portion of T-DNA from Left border to Right border is shown in this figure. Gene of interest (OsGolS1/2; pre-mature/mature) was placed under the constitutive expression of CaMV 35S promoter and NOS terminator. Hygromycin resistant hptII gene is present in the vector backbone as plant selection and GUS is present as reporter gene. b Stages of callus initiation and Agrobacterium mediated transformation. (A) Dehusked seeds of rice IR64 cultivar kept on callus initiation medium (CIM), (B) Mature scutellar calli after 4 weeks of culture, ready for transformation, (C-D) Selection of infected calli in selection medium (CIM + Hyg-50 mg/L; two rounds, each for 15 days). c Immunoblot showing overexpression of his-tagged OsGolS1& OsGolS2 proteins (pre-mature and mature) in rice calli. Lane1: pre-mature OsGolS1, lane2: mature OsGolS1, lane3: pre-mature OsGolS2 and lane4: mature OsGolS2. Lane(s) 1&3 showed sign of degradation. Distinct bands of ~ 39.5 kDa and ~ 37 kDa full-length functional proteins were detected in lanes 2 and 4, respectively (TIFF 1782 kb)

Online Resource 8

(ESM_8.tif): Quantitative RT-PCR expression of GUS in POsGolS1 and POsGolS2 in 7-day-old etiolated rice seedlings under a salt stress (0, 50 and 150 mM NaCl) and b dehydration stress (0, 10 and 20% PEG) (TIFF 382 kb)

Online Resource 9

(ESM_9.tif): Semi-quantitative (a-b) and quantitative (c-d) PCR for first exons (OsGolS1: 326 bp; OsGolS2: 774 bp). The samples used were one-month old plants grown under normal photoperiod (16:8 light:dark) at control conditions treated with low salt (50 mM NaCl) for 8–11 days; each treated with cycloheximide with an untreated control. DAS—days after stress; D1—drug treatment1; FE—first exon. In drug treatment 1, whole seedlings were immersed in the drug. ‘*’ represent p-value < 0.05 as calculated through student’s t-test (TIFF 1169 kb)

Online Resource 10

(ESM_10.tif): Quantitative RT-PCR from nuclear mRNA of leaves of 10 days old rice seedlings for the detection of NMD-sensitive transcripts (OsGolS1 and OsGolS2) (TIFF 316 kb)

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Mukherjee, S., Sengupta, S., Mukherjee, A. et al. Abiotic stress regulates expression of galactinol synthase genes post-transcriptionally through intron retention in rice. Planta 249, 891–912 (2019). https://doi.org/10.1007/s00425-018-3046-z

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