A phytocyanin-related early nodulin-like gene, BcBCP1, cloned from Boea crassifolia enhances osmotic tolerance in transgenic tobacco

Using the mRNA differential display combined with 5' rapid amplification of cDNA ends (RACE) technique, an early nodulin-like protein gene (BcBCP1) (accession no. AY243047.1) was isolated from drought-treated Boea crassifolia leaves. The full-length cDNA of BcBCP1 consists of 844 bp nucleotides and has an open reading frame of 606 bp, encoding a putative polypeptide of 201 amino acids with a predicted molecular mass of 22 kDa and a pI of 5.13. The putative protein precursor contains four sequence domains, including a 27 amino acid hydrophobic N-terminal transit peptide, a 100 amino acid phytocyanin-homologous globular domain, a 51 amino acid hydroxyproline-rich cell wall structural protein domain, and a 22 amino acid hydrophobic extension domain. Sequence alignment defined the encoded protein as an early nodulin-like protein, and the absence of key ligands implies that it is unlikely to bind copper. BcBCP1 expression was strongly induced by dehydration, salinity and abscisic acid (ABA), slightly induced by moderate heat shock, and weakly inhibited by low temperature, methyl jasmonic acid (MeJA), and a low concentration of salicylic acid (SA). Overexpression of BcBCP1 in tobacco under the control of CaMV 35S promoter enhanced tolerance to osmotic stress, as indicated by the less impaired growth, less damaged membrane integrity and lower lipid peroxidation levels after osmotic stress. Transgenic tobacco lines overexpressing BcBCP1 showed higher photosynthetic rates, higher antioxidant enzyme activities and higher cytosyl ascorbic peroxidase transcription levels than non-transgenic tobacco plants, both under normal conditions and under osmotic stress.