Genetic improvement of Chinese cabbage for salt and drought tolerance by constitutive expression of a B. napus LEA gene

Abstract This is the first report on transformation of Chinese cabbage with late embryogenesis abundant (LEA) gene. Transgenic Chinese cabbage (Brassica campestris ssp. pekinensis) expressing a B. napus late embryogenesis abundant protein gene had been generated. Infection by Agrobacterium strain LBA4404 containing the binary vector pIG121–LEA, which carried LEA protein gene linking to CaMV promoter and terminator sequences, and the neomycin phosphotransferase II (NPTII) gene, was applied. Generated shoots were selected by their growth ability on revised MS medium containing kanamycin sulphate. Transgenic Chinese cabbage plants demonstrated enhanced growth ability under salt- and drought-stress conditions. The increased tolerance was reflected by delayed development of damage symptoms caused by stress. The increased tolerance also showed improved recovery upon the removal of stress condition. These results suggest that the genetic modification of Chinese cabbage by LEA protein gene holds considerable potentiality for crop improvement toward environment-stress tolerance.

[1]  T. Miyamoto,et al.  Introduction of the hiC6 Gene, which Encodes a Homologue of a Late Embryogenesis Abundant (LEA) Protein, Enhances Freezing Tolerance of Yeast , 1999 .

[2]  D. Pental,et al.  Agrobacterium-mediated genetic transformation of oilseed Brassica campestris: Transformation frequency is strongly influenced by the mode of shoot regeneration , 1992, Plant Cell Reports.

[3]  Kunihiko,et al.  Factors Influencing Agrobacterium-mediated Transformation of Brassica rapa L. , 1997 .

[4]  M. Montagu,et al.  Transfection and transformation of Agrobacterium tumefaciens , 1978, Molecular and General Genetics MGG.

[5]  B. Ford-Lloyd,et al.  The effects of acetosyringone and pH on Agrobacterium-mediated transformation vary according to plant species , 1991, Plant Cell Reports.

[6]  R. Kalla,et al.  Agrobacterium tumefaciens‐mediated barley transformation , 1997 .

[7]  M. Moloney,et al.  High efficiency transformation ofBrassica napus usingAgrobacterium vectors , 1989, Plant Cell Reports.

[8]  W. Marcotte,et al.  The wheat LEA protein Em functions as an osmoprotective molecule in Saccharomyces cerevisiae , 2004, Plant Molecular Biology.

[9]  N. Kishimoto,et al.  Sequence variation, differential expression and chromosomal location of rice chitinase genes , 1993, Molecular and General Genetics MGG.

[10]  J. Palta,et al.  Responses to abiotic stresses. , 1998 .

[11]  H. Nguyen,et al.  HVA1, a LEA gene from barley confers dehydration tolerance in transgenic rice (Oryza sativa L.) via cell membrane protection , 2004 .

[12]  L. Dure.  A repeating 11-mer amino acid motif and plant desiccation. , 1993, The Plant journal : for cell and molecular biology.

[13]  Wilhelm Gruissem,et al.  Biochemistry & Molecular Biology of Plants , 2002 .

[14]  K. Okada,et al.  Efficient transformation of Arabidopsis thaliana: comparison of the efficiencies with various organs, plant ecotypes and Agrobacterium strains , 1992, Plant Cell Reports.

[15]  K. Wakui,et al.  Isolation and expression of Lea gene in desiccation-tolerant microspore-derived embryos in Brassica spp. , 2002, Physiologia plantarum.

[16]  Ho,et al.  Improved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene. , 2000, Plant science : an international journal of experimental plant biology.

[17]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[18]  A. Ohta,et al.  A lea-class gene of tomato confers salt and freezing tolerance when expressed in Saccharomyces cerevisiae. , 1996, Gene.

[19]  R. Wu,et al.  Genetic improvement of Basmati rice for salt and drought tolerance by regulated expression of a barley Hva1 cDNA , 2002 .

[20]  G. Scoles,et al.  Development of an efficient Agrobacterium-mediated transformation system for Brassica carinata , 1998, Plant Cell Reports.

[21]  C. Bhatia,et al.  Transgenic plants of mustard Brassica juncea (L.) czern and coss , 1990 .

[22]  M. De Block,et al.  Transformation of Brassica napus and Brassica oleracea Using Agrobacterium tumefaciens and the Expression of the bar and neo Genes in the Transgenic Plants. , 1989, Plant physiology.

[23]  I. Nishiyama,et al.  Effects of Foliar and Root-Applied Benzylaminopurine on Tillering of Rice Plants Grown in Hydroponics , 2001 .

[24]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .

[25]  T. Ho,et al.  Expression of a Late Embryogenesis Abundant Protein Gene, HVA1, from Barley Confers Tolerance to Water Deficit and Salt Stress in Transgenic Rice , 1996, Plant physiology.

[26]  Y. Takahata,et al.  Medium and genotype factors influencing shoot regeneration from cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis) , 1998, Plant Cell Reports.