Advertisement

Journal of Plant Biology

, Volume 50, Issue 3, pp 336–343 | Cite as

CbLEA, a NovelLEA Gene fromChorispora bungeana, Confers Cold Tolerance in Transgenic Tobacco

  • Hua Zhang
  • Rui Xia Zhou
  • Li Jing Zhang
  • Ruo Yu Wang
  • Li Zhe An
Article

Abstract

A novel late embryogenesis abundant (LEA) gene (AY804193), namedCbLEA, has now been isolated fromChorispora bungeana. This rare alpine subnival plant can survive sudden snowstorms and low temperatures. The full-lengthCbLEA is 842 bp, with an open reading frame encoding 169 ami no acids. The putative molecular weight ofCbLEA protein is 17.9 kDa, with an estimatedpl of 6.45. To investigate the functioning of thisCbLEA protein in cold-stress tolerance,CbLEA was introduced into tobacco under the control of the CaMV35S promoter. Second-generation (R1) transgenic tobacco plants exhibited significantly increased tolerance to cold. These transgenics maintained lower malondialdehyde (MDA) contents and electrolyte leakage (EL) but their relative water content (RWC) was significantly higher compared with non-transgenic plants under chilling stress. Further experimental results showed that non-transgenic plants had severe freezing damage after exposure to -2°C for 1 h, whereas the transgenics suffered only slight injury under the same conditions. Moreover, survival was longer in the latter genotype at that temperature. The extent of increased cold tolerance was positive correlated with the level ofCbLEA protein accumulation, and was also reflected by the delayed development of damage symptoms. This indicates thatCbLEA is an excellent stress tolerance gene, and holds considerable potential as a new molecular tool for engineering improved plant genetics.

Keywords

chilling stress Chorispora bungeana cold tolerance freezing stress LEA gene transgenic tobacco 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Literature ctied

  1. An LZ, Liu YH, Feng HY, Feng GN, Chen GD (2000) Studies on characteristics of element contents of altifrigetic subnival vegetation at the source area of Urumqi River. Acta Bot Boreal-Occident Sin20: 80–86Google Scholar
  2. Ayi tu R, Tan DY, Li ZJ, Yao F (1998) The relationship between the structures of vegetative organs in C.bungeana and its environment. J Xinjiang Agric Univ21: 273–277Google Scholar
  3. Babu RC, Zhang J, Blum A, Ho DTH, Wu R, Nguyen HT (2004)HVA1, a LEA gene from barley, confers dehydration tolerance in transgenic rice(Oryza sativa L) via cell membrane protection. Plant Sci166: 855–862CrossRefGoogle Scholar
  4. Baker J, Steel C, Dure L III (1988) Sequence and characterization of 6 Lea proteins and their genes from cotton. Plant Mol Biol11: 277–291CrossRefGoogle Scholar
  5. Bradford M (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem72: 248–254PubMedCrossRefGoogle Scholar
  6. Danyluk J, Perron A, Houde M, Limin A, Fowler B, Benhamou N, Sarhan F (1998) Accumulation of an acidic dehydrin in the vicinity of the plasma membrane during cold acclimation of wheat. Plant Cell10: 623–638PubMedCrossRefGoogle Scholar
  7. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus12: 13–15Google Scholar
  8. Dure L III (1993) A repeating 11-mer amino acid motif and plant desiccation. Plant J3: 363–369PubMedCrossRefGoogle Scholar
  9. Dure L III,Croch M, Harada J, Ho DTH, Mundy J, Quatrano RS, Thomas T, Sung ZR (1989) Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol Biol12: 475–486CrossRefGoogle Scholar
  10. Dure L III,Greenway SC, Galau GA (1981) Developmental biochemistry of cottonseed embryogenesis and germination: Changing messenger ribonucleic acid populations as shown byin vitro andin vivo protein synthesis. Biochemistry20: 4162–4168PubMedCrossRefGoogle Scholar
  11. Fu XY, Chang JF, An LZ, Zhang MX, Xu SJ, Chen T, Liu YH, Xin H,Wang JH (2006) Association of the cold-hardiness ofChorispora bungeana with the distribution and accumulation of calcium in the cells and tissues. Environ Exp Bot55: 282–293CrossRefGoogle Scholar
  12. Hasegawa M, Bressan R, Zhu JK, Bohnert HJ (2000) Plant cellular and molecular responses to high salinity. Annu Rev Plant Physiol Plant Mol Biol51: 463–469PubMedCrossRefGoogle Scholar
  13. Hofgen R, Willmitzer L (1988) Storage of competent cells forAgrobacterium transformation. Nucl Acids Res16: 9877PubMedCrossRefGoogle Scholar
  14. Honjoh Kl, Oda Y, Takata R, Miyamoto T, Hatano S (1999) Introduction of the hiC6 gene, which encodes a homologue of a late embryogenesis abundant (LEA) protein, enhances freezing tolerance of yeast. J Plant Physiol155: 509–512Google Scholar
  15. Imai R, Chang L, Ohta A, Bray EA, Takagi M (1996) A lea-class gene of tomato confers salt and freezing tolerance when expressed inSaccharomyces cerevisiae. Gene170: 243–248PubMedCrossRefGoogle Scholar
  16. Jaakola L, Pirtila AM, Halonen M, Hohtola A (2001) Isolation of high quality RNA from bilberry(Vaccinium myrtillus L.) fruit. Mol Biotech19: 203–210CrossRefGoogle Scholar
  17. Jun SS, Yang JY, Choi HJ, Kim NR, Park MC, Hong YN (2005) Altered physiology in trehalose-producing transgenic tobacco plants: Enhanced tolerance to drought and salinity stresses. J Plant Biol48: 456–466CrossRefGoogle Scholar
  18. Kim HS, Lee JH, Kim JJ, Kim CH, Jun SS, Hong YN (2005) Molecular and functional characterization ofCaLEA6, the gene for a hydrophobic LEA protein fromCapsicum annuum. Gene344: 115–123PubMedCrossRefGoogle Scholar
  19. Kim SH, Lee HS, Song WY, Choi KS, Hur Y (2007) Chloroplast-tar-geted BrMT1 (Brassica rapa type-1 metallothionein) enhances resistance to cadmium and ROS in transgenicArabidopsis plants. J Plant Biol50: 1–7CrossRefGoogle Scholar
  20. Liang CY, Xi Y, Shu J, Li J, Yang JL, Che KP, Jin DM, Liu XL, Weng ML, He YK, Wang B (2004) Construction of a BAC library ofPhyscomitrella patens and isolation of aLEA gene. Plant Sci167: 491–498CrossRefGoogle Scholar
  21. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, New York, pp 440Google Scholar
  22. Moons A, de KA, van MM (1997) A group 3LEA cDNA of rice, responsive to abscisic acid, but not to jasmonic acid, shows variety-specific differences in salt stress response. Gene191: 197–204PubMedCrossRefGoogle Scholar
  23. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant15: 473–497CrossRefGoogle Scholar
  24. Park BJ, Liu ZC, Kanno A, Kameya T (2005) Genetic improvement of Chinese cabbage for salt and drought tolerance by constitutive expression of aB. napus LEA gene. Plant Sci169: 553–558CrossRefGoogle Scholar
  25. Ramanjulu S, Bartels D (2002) Drought- and desiccation-induced modulation of gene expression in plants. Plant Cell Environ25: 141–151PubMedCrossRefGoogle Scholar
  26. Raynal M, Gaubier P, Grellet F, Delseny M (1990) Nucleotide sequence of a radish cDNA clone coding for a late embryo-genesis abundant (LEA) protein. Nucl Acids Res18: 6132PubMedCrossRefGoogle Scholar
  27. Rohila JS, Jain RK, Wu R (2002) Genetic improvement of Basmati rice for salt and drought tolerance by regulated expression of a barleyHva1 cDNA. Plant Sci163: 525–532CrossRefGoogle Scholar
  28. Seki M, Narusaka M, Abe H, Kasuga M, Yamaguchi-Shinozaki K, Carninci P, Hayashizaki Y, Shinozaki K (2001) Monitoring the expression pattern of 1300Arabidopsis genes under drought and cold stresses by using a full-length cDNA microarray. Plant Cell13: 61–72PubMedCrossRefGoogle Scholar
  29. Shih MD, Lin SC, Hsieh JS, Tsou CH, Chow TY, Lin TP, Hsing Yl (2004) Gene cloning and characterization of a soybean(Glycine max L) LEA protein, GmPM16. Plant Mol Biol56: 689–703PubMedCrossRefGoogle Scholar
  30. Sivamani E, Bahieldinl 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 barleyHVA1 gene. Plant Sci155: 1–9PubMedCrossRefGoogle Scholar
  31. Thomashow MF (1998) Role of cold-responsive genes in plant freezing tolerance. Plant Physiol118: 1–7PubMedCrossRefGoogle Scholar
  32. Walker K, Croteau R (2000) Taxol biosynthesis-molecular cloning of a benzoyl-CoA-taxane 2a-O-benzoyltransferase cDNA from Taxus and functional expression inEscherichia coli. Proc Natl Acad Sci USA97: 13591–13596PubMedCrossRefGoogle Scholar
  33. Wang YC, Jiang J, Zhao X, Liu GF, Yang CR, Zhan LP (2006) A novelLEA gene fromTamarix androssowii confers drought tolerance in transgenic tobacco. Plant Sci171: 655–666CrossRefGoogle Scholar
  34. Weatherley PE (1950) Studies in the water relations of cotton plants: The field measurement of water deficit in leaves. New Phytol49: 81–87CrossRefGoogle Scholar
  35. Xu D, Duan X, Wang B, Hong B, Ho TH, 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 Physiol110: 249–257PubMedGoogle Scholar

Copyright information

© The Botanical Society of Korea 2007

Authors and Affiliations

  1. 1.Key Laboratory of Arid and Grassland Agroecology of Ministry of Education, School of Life SciencesLanzhou UniversityLanzhouChina
  2. 2.College of Pastoral Agriculture Science and TechnologyLanzhou UniversityLanzhouChina
  3. 3.Cold and Arid Regions Environmental and Engineering Research InstituteChinese Academy of SciencesLanzhouChina

Personalised recommendations