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Over-expression of a LEA gene in rice improves drought resistance under the field conditions

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Abstract

Late embryogenesis abundant (LEA) proteins have been implicated in many stress responses of plants. In this report, a LEA protein gene OsLEA3-1 was identified and over-expressed in rice to test the drought resistance of transgenic lines under the field conditions. OsLEA3-1 is induced by drought, salt and abscisic acid (ABA), but not by cold stress. The promoter of OsLEA3-1 isolated from the upland rice IRAT109 exhibits strong activity under drought- and salt-stress conditions. Three expression constructs consisting of the full-length cDNA driven by the drought-inducible promoter of OsLEA3-1 (OsLEA3-H), the CaMV 35S promoter (OsLEA3-S), and the rice Actin1 promoter (OsLEA3-A) were transformed into the drought-sensitive japonica rice Zhonghua 11. Drought resistance pre-screening of T1 families at anthesis stage revealed that the over-expressing families with OsLEA3-S and OsLEA3-H constructs had significantly higher relative yield (yield under drought stress treatment/yield under normal growth conditions) than the wild type under drought stress conditions, although a yield penalty existed in T1 families under normal growth conditions. Nine homozygous families, exhibiting over-expression of a single-copy of the transgene and relatively low yield penalty in the T1 generation, were tested in the field for drought resistance in the T2 and T3 generations and in the PVC pipes for drought tolerance in the T2 generation. Except for two families (transformed with OsLEA3-A), all the other families (transformed with OsLEA3-S and OsLEA3-H constructs) had higher grain yield than the wild type under drought stress in both the field and the PVC pipes conditions. No significant yield penalty was detected for these T2 and T3 families. These results indicate that transgenic rice with significantly enhanced drought resistance and without yield penalty can be generated by over-expressing OsLEA3-1 gene with appropriate promoters and following a bipartite (stress and non-stress) in-field screening protocol.

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References

  • Bake J, Steele C, Dure LI (1988) Sequence and characterization of 6 LEA proteins and their genes from cotton. Plant Mol Biol 11:277–291

    Article  Google Scholar 

  • Baker SS, Wilhelm KS, Thomashow MF (1994) The 5′-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold-, drought- and ABA-regulated gene expression. Plant Mol Biol 24:701–713

    Article  PubMed  CAS  Google Scholar 

  • Bray EA, Bailey-Serres J, Weretilnyk E (2000) Responses to abiotic stresses. In: Gruissem W, Buchannan B, Jones R (eds) Biochemistry and molecular biology of plants. American Society of Plant Physiologists, Rockville, pp 1158–1249

    Google Scholar 

  • Chen H, Tang W, Xu C, Li X, Lin Y, Zhang Q (2005) Transgenic indica rice plants harboring a synthetic cry2A* gene of Bacillus thuringiensis exhibit enhanced resistance against lepidopteran rice pests. Theor Appl Genet 111:1330–1337

    Article  PubMed  CAS  Google Scholar 

  • Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG, Thompson JD (2003) Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31:3497–3500

    Article  PubMed  CAS  Google Scholar 

  • Chu ZH, Peng KM, Zhang LD, Zhou B, Wei J, Wang SP (2003) Construction and characterization of a normalized whole-life-cycle cDNA library of rice. Chin Sci Bull 48:229–235

    Article  CAS  Google Scholar 

  • Cuming AC (1999) LEA proteins. In: Shewry PR, Casey R (eds) Seed proteins. Kluwer, Dordrecht, pp 753–780

    Google Scholar 

  • Curry J, Walker-Simmons MK (1993) Unusual sequence of group 3 LEA (II) mRNA inducible by dehydration stress in wheat. Plant Mol Biol 21:907–912

    Article  PubMed  CAS  Google Scholar 

  • Curry J, Morris CF, Walker-Simmons MK (1991) Sequence analysis of a cDNA encoding a group 3 LEA mRNA inducible by ABA or dehydration stress in wheat. Plant Mol Biol 16:1073–1076

    Article  PubMed  CAS  Google Scholar 

  • Dubouzet JG, Sakuma Y, Ito Y, Kasuga M, Dubouzet EG, Miura S, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J 33:751–763

    Article  PubMed  CAS  Google Scholar 

  • Dure LIII, Crouch M, Harada J, Ho T-H, Mundy J, Quatrano RS, Thomas T, Sung ZR (1989) Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol Biol 12:475–486

    Article  CAS  Google Scholar 

  • Garay-Arroyo A, Colmenero-Flores JM, Garciarrubio A, Covarrubias AA (2000) Highly hydrophilic proteins in prokaryotes and eukaryotes are common during conditions of water deficit. J Biol Chem 275:5668–5674

    Article  PubMed  CAS  Google Scholar 

  • Garg AK, Kim JK, Owens TG, Ranwala AP, Choi YD, Kochian LV, Wu RJ (2002) Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses. Proc Natl Acad Sci USA 99:15898–15903

    Article  PubMed  CAS  Google Scholar 

  • Goyal K, Walton LJ, Tunnacliffe A (2005) LEA proteins prevent protein aggregation due to water stress. Biochem J 388:151–157

    Article  PubMed  CAS  Google Scholar 

  • Grelet J, Benamar A, Teyssier E, Avelange-Macherel MH, Grunwald D, Macherel D (2005) Identification in pea seed mitochondria of a late-embryogenesis abundant protein able to protect enzymes from drying. Plant Physiol 137:157–167

    Article  PubMed  CAS  Google Scholar 

  • Hayashi H, Alia, Mustardy L, Deshnium P, Ida M, Murata N (1997) Transformation of Arabidopsis thaliana with the codA gene for choline oxidase; accumulation of glycinebetaine and enhanced tolerance to salt and cold stress. Plant J 12:133–142

    Article  PubMed  CAS  Google Scholar 

  • Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271–282

    Article  PubMed  CAS  Google Scholar 

  • Hoisington D, Jiang C, Khairallah M, Ribaut JM, Bohn M, Melchinger A, Willcox M, Gonzalez-de-Leon D (1996) QTL for insect resistance and drought tolerance in tropical maize: prospects for marker assisted selection. Symp Soc Exp Biol 50:39–44

    PubMed  CAS  Google Scholar 

  • Holmström K-O, Mäntylä E, Welin B, Mandal A, Palva ET, Tunnela OE, Londesborough J (1996) Drought tolerance in tobacco. Nature 379:683–684

    Article  Google Scholar 

  • Hong B, Uknes SJ, Ho T-HD (1988) Cloning and characterization of a cDNA encoding a mRNA rapidly induced by ABA in barley aleurone layers. Plant Mol Biol 11:495–506

    Article  CAS  Google Scholar 

  • Hu H, Dai M, Yao J, Xiao B, Li X, Zhang Q, Xiong L (2006) Overexpressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice. Proc Natl Acad Sci USA 103:12987–12992

    Article  PubMed  CAS  Google Scholar 

  • Ingram J, Bartels D (1996) The molecular basis of dehydration tolerance in plants. Annu Rev Plant Physiol Plant Mol Biol 47:377–403

    Article  PubMed  CAS  Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    PubMed  CAS  Google Scholar 

  • Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1999) Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol 17:287–291

    Article  PubMed  CAS  Google Scholar 

  • Kim HS, Lee JH, Kim JJ, Kim CH, Jun SS, Hong YN (2005) Molecular and functional characterization of CaLEA6, the gene for a hydrophobic LEA protein from Capsicum annuum. Gene 344:115–123

    Article  PubMed  CAS  Google Scholar 

  • Kishor P, Hong Z, Miao GH, Hu C, Verma D (1995) Over-expression of Δ-pyrroline-5-carboxylate synthetase increases proline production and confers osmotolerance in transgenic plants. Plant Physiol 108:1387–1394

    PubMed  CAS  Google Scholar 

  • Lin YJ, Zhang Q (2005) Optimising the tissue culture conditions for high efficiency transformation of indica rice. Plant Cell Rep 23:540–547

    Article  PubMed  CAS  Google Scholar 

  • Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1998) Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression, respectively, in Arabidopsis. Plant Cell 10:1391–1406

    Article  PubMed  CAS  Google Scholar 

  • Maqbool B, Zhong H, El-Maghraby Y, Ahmad A, Chai B, Wang W, Sabzikar R, Sticklen B (2002) Competence of oat (Avena sativa L.) shoot apical meristems for integrative transformation, inherited expression, and osmotic tolerance of transgenic lines containing hva1. Theor Appl Genet 105:201–208

    Article  PubMed  CAS  Google Scholar 

  • McElroy D, Zhang W, Cao J, Wu R (1990) Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2:163–171

    Article  PubMed  CAS  Google Scholar 

  • Meyer P (2000) Transcriptional transgene silencing and chromatin components. Plant Mol Biol 43:221–234

    Article  PubMed  CAS  Google Scholar 

  • Moons A, Bauw G, Prinsen E, Van Montagu M, Van der Straeten D (1995) Molecular and physiological responses to abscisic acid and salts in roots of salt-sensitive and salt-tolerant indica rice varieties. Plant Physiol 107:177–186

    Article  PubMed  CAS  Google Scholar 

  • Moons A, De Keyser A, Van Montagu M (1997) A group 3 LEA cDNA of rice, responsive to abscisic acid, but not to jasmonic acid, shows variety-specific differences in salt stress response. Gene 191:197–204

    Article  PubMed  CAS  Google Scholar 

  • NDong C, Danyluk J, Wilson KE, Pocock T, Huner NP, Sarhan F (2002) Cold-regulated cereal chloroplast late embryogenesis abundant-like proteins: molecular characterization and functional analyses. Plant Physiol 129:1368–1381

    Article  PubMed  CAS  Google Scholar 

  • Park S, Li J, Pittman JK, Berkowitz GA, Yang H, Undurraga S, Morris J, Hirschi KD, Gaxiola RA (2005) Up-regulation of a H+-pyrophosphatase (H+-P Pase) as a strategy to engineer drought-resistant crop plants. Proc Natl Acad Sci USA 102:18830–18835

    Article  PubMed  CAS  Google Scholar 

  • Raynal M, Guilleminot J, Gueguen C, Cooke R, Delseny M, Gruber V (1999) Structure, organization and expression of two closely related novel Lea (late-embryogenesis-abundant) genes in Arabidopsis thaliana. Plant Mol Biol 40:153–165

    Article  PubMed  CAS  Google Scholar 

  • Ried JL, Walker-Simmons MK (1993) Group 3 late embryogenesis abundant proteins in desication-tolerant seedlings of wheat (Triticum aestivum L.). Plant Physiol 102:125–131

    PubMed  CAS  Google Scholar 

  • Romero C, Belles J, Vaya J, Serrano R, Culianez-Macia F (1997) Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance. Planta 201:293–297

    Article  CAS  PubMed  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor, New York

    Google Scholar 

  • Shao HB, Liang ZS, Shao MA (2005) LEA proteins in higher plants: structure, function, gene expression and regulation. Colloids Surf B Biointerfaces 45:131–135

    Article  CAS  Google Scholar 

  • Shen B, Jensen RG, Bohnert HJ (1997) Increased resistance to oxidative stress in transgenic plants by targeting mannitol biosynthesis to chloroplasts. Plant Physiol 113:1177–1183

    Article  PubMed  CAS  Google Scholar 

  • Soltis DE, Soltis PS (2003) The role of phylogenetics in comparative genetics. Plant Physiol 132:1790–1800

    Article  PubMed  CAS  Google Scholar 

  • Stam M, Mol JN, Kooter JM (1997) The silence of genes in transgenic plants. Ann Bot 79:3–12

    Article  CAS  Google Scholar 

  • Stempak JM, Sohn KJ, Chiang EP, Shane B, Kim YI (2005) Cell and stage of transformation-specific effects of folate deficiency on methionine cycle intermediates and DNA methylation in an in vitro model. Carcinogenesis 26:981–990

    Article  PubMed  CAS  Google Scholar 

  • Su J, Shen Q, David Ho TH, Wu R (1998) Dehydration-stress-regulated transgene expression in stably transformed rice plants. Plant Physiol 117:913–922

    Article  PubMed  CAS  Google Scholar 

  • Thomashow MF (1998) Role of cold-responsive genes in plant freezing tolerance. Plant Physiol 118:1–8

    Article  PubMed  CAS  Google Scholar 

  • Thomashow MF (1999) Plant Cold Acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599

    Article  PubMed  CAS  Google Scholar 

  • Turner NC (1979) Drought resistance and adaptation to water deficits in crop plants. In: Mussall H (ed) Stress physiology in crop plants, Wiley, New York, pp 343–372

    Google Scholar 

  • Turner NC (1997) Further progress in crop water relations. Adv Agron 58:293–339

    Article  Google Scholar 

  • Wang H, Cutler AJ (1995) Promoters from kin1 and cor6.6, two Arabidopsis thaliana low-temperature- and ABA-inducible genes, direct strong β-glucuronidase expression in guard cells, pollen and young developing seeds. Plant Mol Biol 28:619–634

    Article  PubMed  CAS  Google Scholar 

  • Wang H, Datla R, Georges F, Loewen M, Cutler AJ (1995) Promoters from kin1 and cor6.6, two homologous Arabidopsis thaliana genes: transcriptional regulation and gene expression induced by low temperature, ABA, osmoticum and dehydration. Plant Mol Biol 28:605–617

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Ying J, Kuzma M, Chalifoux M, Sample A, McArthur C, Uchacz T, Sarvas C, Wan J, Dennis DT, McCourt P, Huang Y (2005) Molecular tailoring of farnesylation for plant drought tolerance and yield protection. Plant J 43:413–424

    Article  PubMed  CAS  Google Scholar 

  • Wise (2003) Leaping to conclusions: a computational reanalysis of late embryogenesis abundant proteins and their possible roles. BMC Bioinformatics 4:52

    Article  PubMed  Google Scholar 

  • Wu C, Li X, Yuan W, Chen G, Kilian A, Li J, Xu C, Zhou DX, Wang S, Zhang Q (2003) Development of enhancer trap lines for functional analysis of the rice genome. Plant J 35:418–427

    Article  PubMed  CAS  Google Scholar 

  • Xiong L, Yang Y (2003) Disease resistance and abiotic stress tolerance in rice are inversely modulated by an abscisic acid-inducible mitogen-activated protein kinase. Plant Cell 15:745–759

    Article  PubMed  CAS  Google Scholar 

  • Xiong L, Schumaker KS, Zhu JK (2002) Cell signaling during cold, drought, and salt stress. Plant Cell 14(Suppl):S165–S183

    PubMed  CAS  Google Scholar 

  • Xu D, Duan X, Wang B, Hong B, Ho T, Wu R (1996) Expression of a late embryogenesis abundant protein gene, HVA1, from barley confers tolerance to water deficit and salt stress in transgenic rice. Plant Physiol 110:249–257

    PubMed  CAS  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (1994) A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. Plant Cell 6:251–264

    Article  PubMed  CAS  Google Scholar 

  • Yoon DB, Kang KH, Kim HJ, Ju HG, Kwon SJ, Suh JP, Jeong OY, Ahn SN (2006) Mapping quantitative trait loci for yield components and morphological traits in an advanced backcross population between Oryza grandiglumis and the O. sativa japonica cultivar Hwaseongbyeo. Theor Appl Genet 112:1052–1062

    Article  PubMed  CAS  Google Scholar 

  • Yue B, Xue W, Xiong L, Yu X, Luo L, Cui K, Jin D, Xing Y, Zhang Q (2006) Genetic basis of drought resistance at reproductive stage in rice: separation of drought tolerance from drought avoidance. Genetics 172:1213–1228

    Article  PubMed  Google Scholar 

  • Zhang HX, Hodson JN, Williams JP, Blumwald E (2001) Engineering salt-tolerant Brassica plants: characterization of yield and seed oil quality in transgenic plants with increased vacuolar sodium accumulation. Proc Natl Acad Sci USA 98:12832–12836

    Article  PubMed  CAS  Google Scholar 

  • Zhang Q, Saghai Maroof MA, Lu TY, Shen BZ (1992) Genetic diversity and differentiation of indica and japonica rice detected by RFLP analysis. Theor Appl Genet 83:495–499

    Google Scholar 

  • Zhang X, Zhou S, Fu Y, Su Z, Wang X, Sun C (2006) Identification of a drought tolerant introgression line derived from Dongxiang common wild rice (O. rufipogon Griff.). Plant Mol Biol 62:247–259

    Article  PubMed  CAS  Google Scholar 

  • Zhao HW, Chen YJ, Hu YL, Gao Y, Lin ZP (2000a) Construction of a trehalose-6-phosphate synthase gene driven by drought-responsive promoter and expression of drought-resistance in transgenic tobacco. Acta Bot Sin 42:616–619

    CAS  Google Scholar 

  • Zhao SH, Wang FZ, Lu L, Zhang HY, Zhang XY (2000b) Breeding and selection of drought resistant and salt tolerant wheat variety Cang 6001. Acta Agric Boreall Sin 15:113–117

    Google Scholar 

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Acknowledgments

We thank Drs. Abraham Blum and Qifa Zhang for suggestions on drought resistance testing protocol of rice in the field. This research was supported by the National Program on Basic Research and Development, the National Special Key Project on Functional Genomics of Rice, the National Natural Science Foundation of China, and the Rockefeller Foundation.

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  1. National Center of Plant Gene Research (Wuhan), National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China

    Benze Xiao, Yuemin Huang, Ning Tang & Lizhong Xiong

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  1. Benze Xiao
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  2. Yuemin Huang
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Correspondence to Lizhong Xiong.

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Xiao, B., Huang, Y., Tang, N. et al. Over-expression of a LEA gene in rice improves drought resistance under the field conditions. Theor Appl Genet 115, 35–46 (2007). https://doi.org/10.1007/s00122-007-0538-9

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