Abstract
Drought is a major agricultural menace reducing crop productivity and limiting the successful realization of land potential throughout the world. Therefore, breeding common wheat with improved drought-tolerance via genetic manipulation is of great importance. We have introduced the betA gene encoding choline dehydrogenase from Escherichia coli into common wheat (Triticum aestivum L.) by Agrobacterium-mediated transformation. Various levels of expression of the betA gene were confirmed by RT-PCR among the transgenic lines and different levels of glycine betaine accumulation were detected in these lines. Several wheat transgenic lines with different betA expression levels in the T3 generation and wild-type (WT) were selected to test their performance under drought stress conditions. Water deficit in plants caused a reduction in photosynthesis and activity of the PSII complex and resulted in increased accumulation of osmolytes. Drought stress also led to lower membrane stability along with much higher activities of superoxide dismutase and peroxidase in all wheat lines. However, wheat lines that were transgenic for the betA gene were less injured and exhibited greater root length and growth compared with the WT. It was concluded that the amount of injury to the wheat plants was negatively correlated with the level of accumulation of glycine betaine, and the glycine betaine acted as an important osmoprotectant in transgenic plants to improve root growth, and enhance the resistance of transgenic plants to drought stress.
Similar content being viewed by others
Abbreviations
- BADH:
-
Betaine aldehyde dehydrogenase
- CDH:
-
Choline dehydrogenase
- CMO:
-
Choline monooxygenase
- GB:
-
Glycine betaine
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
- MDA:
-
Malondialdehyde
- RWC:
-
Relative water content
- Ls:
-
Stomatal limitation value
- NBT:
-
Nitroblue tetrazolium
- POD:
-
Peroxidase
- SOD:
-
Superoxide dismutase
- ROS:
-
Reactive oxygen species
- WT:
-
Wild-type
References
Abebe T, Guenzi AC, Martin B, Cushman JC (2003) Tolerance of mannitol-accumulating transgenic wheat to water stress and salinity. Plant Physiol 131:1748–1755
Allakhverdiev SI, Feyziev YM, Ahemd A, Hayashi H, Aliev JA, Klimov VV, Murata N, Carpentier R (1996) Stabilization of oxygen evolution and primary electron transport reactions in photosystemII against heat stress with glycinebetaine and sucrose. J Photochem Photobiol 34:149–157
Allard F, Houde M, Kröl M, Ivanov A, Huner NPA, Sarhan F (1998) Betaine improves freezing tolerance in wheat. Plant Cell Physiol 39:1194–1202
Arnon DI (1949) Copper enzymes in isolated chloroplasts: polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207
Berry JA, Downton WJS (1982) Environmental regulation of photosynthesis. In: Govind J (ed) Photosynthesis (Vol II). Academic Press, New York, pp 263–345
Bradford M (1976) A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Carillo P, Mastrolonardo G, Nacca F, Parisi D, Verlotta A, Fuggi A (2008) Nitrogen metabolism in durum wheat under salinity: accumulation of proline and glycine betaine. Funct Plant Bio 35:412–426
Cornic G, Massacci A (1996) Leaf photosynthesis under drought stress. In: Baker NR (ed) Photosynthesis and the environment: advance in photosynthesis research. Kluwer, Dordrecht, p 347
Fan L, Zheng S, Wang X (1997) Antisense suppression of phospholipase Dα retards abscisic acid- and ethylene promoted senescence of postharvest arabidopsis leaves. Plant Cell 9:2183–2196
Gaxiola RA, Li J, Undurraga S, Dang LM, Allen GJ, Alper SL, Fink GR (2001) Drought- and salt-tolerant plants result from overexpression of the AVP1 H+-pump. Proc Natl Acad Sci USA 98:11444–11449
Giannopolitis CN, Ries SK (1977) Superoxide dismutase. I. Occurrence in higher plants. Plant Physiol 59:309–314
Li GS, Yang AF, Zhang JR, Bi YP, Shan L (2000) Genetic transformation of calli from maize and regeneration of herbicide-resistant plantlets. Chinese Sci Bull 46:563–565
Li Y, Wang W, Yang X, Zou Q (2005) Expression of BADH and betaine content in wheat cultivars with different drought resistance under drought stress. Acta Agron Sin 31:425–430
Lilius G, Holmberg N, Bulow L (1996) Enhanced NaCl stress tolerance in transgenic tobacco expressing bacterial choline dehydrogenase. BioTechnol 14:177–180
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
Lv SL, Yang AF, Zhang KW, Wang L, Zhang J (2007) Increase of glycinebetaine synthesis improves drought tolerance in cotton. Mol Breed 20:233–248
Ma QQ, Wang W, Li YH, Li DQ, Zou Q (2006) Alleviation of photoinhibition in drought-stressed wheat (Triticum aestivum) by foliar-applied glycinebetaine. J plant physiol 163(2):165–175
Mamedov M, Hayashi H, Murata N (1993) Effects of glycinebetaine and unsaturation of membrane lipids on heat stability of photosynthetic electron-transport and phosphorylation reactions in Synechocystis PCC6803. Biochim Biophys Acta 1142:1–5
Mandhania S, Madan S, Sawhney V (2006) Antioxidant defense mechanism under salt stress in wheat seedlings. Biol Plant 50(2):227–231
Mansour MMF (1998) Protection of plasma membrane of onion epidermal cells by glycinebetaine and proline against NaCl stress. Plant Physiol Biochem 36:767–772
Nanjo T, Kobayashi M, Yoshiba Y, Kakubari Y, Yamaguchi-Shinozaki K, Shinozaki K (1999) Antisense suppression of proline degradation improves tolerance to freezing and salinity in Arabidopsis thaliana. FEBS Letters 461:205–210
Ozturk ZN, Talame′ V, Deyholos M, Michalowski CB, Galbraith DW, Gozukirmizi N, Tuberosa R, Bohnert HJ (2002) Monitoring large-scale changes in transcript abundance in drought- and salt-stressed barley. Plant Mol Biol 48:551–573
Passioura J (2007) The drought environment: physical, biological and agricultural perspectives. J Exp Bot 58(22):113–117
Pastori GM, Foyer CH (2002) Common components, networks, and pathways of cross-tolerance to stress: the central role of “redox” and abscisic acid-mediated controls. Plant Physiol 129:460–468
Quan RD, Shang M, Zhang H, Zhao YX, Zhang JR (2004a) Improved chilling tolerance by transformation with betA gene for the enhancement of glycinebetaine synthesis in maize. Plant Sci 166:141–149
Quan RD, Shang M, Zhang H, Zhao YX, Zhang JR (2004b) Engineering of enhanced glycine betaine synthesis improves drought tolerance in maize. Plant Biotechnol J 2:477–486
Rathinasabapathi B, Burnet M, Russell BL, Gage DA, Liao PC, Nye GJ, Scott P, Golbeck JH, Hanson AD (1997) Choline monooxygenase, an unusual iron-sulfur enzyme catalyzing the first step of glycine betaine synthesis in plants: prosthetic group characterization and cDNA cloning. Proc Natl Acad Sci USA 94:3454–3458
Rhodes D, Hanson AD (1993) Quaternary ammonium and tertiary sulfonium compounds in higher plants. Annu Rev Plant Physiol Mol Biol 44:357–384
Sakamoto A, Murata N (1998) Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold. Plant Mol Biol 38:1011–1019
Sakamoto A, Murata N (2002) The role of glycine betaine in the protection of plants from stress: clues from transgenic plants. Plant Cell Environ 25:163–171
Sakamoto A, Valverde R, Alia Chen THH, Murata N (2000) Transformation of arabidopsis with the codA gene for choline oxidase enhances freezing tolerance of plants. Plant J 22:449–453
Saneoka H, Nagasaka C, Hahn DT, Yang W-J, Premachandra GS, Joly RJ, Rhodes D (1995) Salt tolerance of glycinebetaine-deficient and -containing maize lines. Plant Physiol 107:631–638
Saneoka H, Moghaieb REA, Premachandra GS, Fujita K (2004) Nitrogen nutrition and water stress effects on cell membrane stability and leaf water relations in Agrostis palustris Huds. Environ Exp Bot 52:131–138
Sivamani E, Bahieldin A, Wraith JM, Al-Niemi T, Dyer WE, Ho TD, Qu R (2000) Improved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene. Plant Sci 155:1–9
Waditee R, Bhuiyan MNH, Rai V, Aoki K, Tanaka Y, Hibino T, Suzuki S, Takano J, Jagendorf AT, Takabe T, Takabe T (2005) Genes for direct methylation of glycine provide high levels of glycinebetaine and abiotic-stress tolerance in Synechococcus and Arabidopsis. Proc Natl Acad Sci USA 102:1318–1323
Xue ZY, Zhi DY, Xue GP, Zhang H, Zhao YX, Xia GM (2004) Enhanced salt tolerance of transgenic wheat (Tritivum aestivum L.) expressing a vacuolar Na+/H+ antiporter gene with improved grain yields in saline soils in the field and a reduced level of leaf Na+. Plant Sci 167:849–859
Yang X, Liang Z, Lu C (2005) Genetic engineering of the biosynthesis of glycinebetaine enhances photosynthesis against high temperature stress in transgenic tobacco plants. Plant Physiol Preview 138:2299–2309
Yang X, Wen X, Gong H, Lu Q, Yang Z, Tang Y, Liang Z, Lu C (2007) Genetic engineering of the biosynthesis of glycinebetaine enhances thermo tolerance of photosystem II in tobacco plants. Planta 225(3):719–733
Yang X, Liang Z, Wen X, Lu C (2008) Genetic engineering of the biosynthesis of glycinebetaine leads to increased tolerance of photosynthesis to salt stress in transgenic tobacco plants. Plant Mol Biol 66:73–86
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
Zheng Y, Jia A, Ning T, Xu J, Li Z, Jiang G (2008) Potassium nitrate application alleviates sodium chloride stress in winter wheat cultivars differing in salt tolerance. J Plant Physiol 165:1455–1465
Acknowledgments
We are grateful to Dr. Yongbin Yan (Tsinghua University) for their assistance with NMR. This research was supported by Hi-Tech Research and Development (863) Program of China (2007AA10Z175).
We thank Dr. Roberta Greenwood for her help in editing this manuscript.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
He, C., Zhang, W., Gao, Q. et al. Enhancement of drought resistance and biomass by increasing the amount of glycine betaine in wheat seedlings. Euphytica 177, 151–167 (2011). https://doi.org/10.1007/s10681-010-0263-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10681-010-0263-3