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Copper chaperone (CCH) is upregulated during  Arabidopsis  ( Arabidopsis thaliana  L. Columbia) leaf senescence, suggesting that it mobilizes certain metal ions in leaves and transports them to other growing parts of the plants. The CCHs are also involved in defense mechanisms against oxidative stress in  Arabidopsis  and tomato ( Lycopersicon esculentum  Mill. cv. ‘Ailsa Craig’). To elucidate the functions of CCH in poplar, we cloned a  CCH  gene ( PoCCH ) from  Populus  ( Populus alba  ×  P. tremula  var.  glandulosa ) suspension cells and tested its expression in response to various treatments including heavy metals, plant growth regulators and abiotic stresses. The  PoCCH  cDNA is 540 bp in length, including a 55-bp 5′ noncoding domain, a 258-bp open reading frame (ORF), and 227-bp 3′ termination region. The coding region of  PoCCH  represents a putative 85-amino-acid protein with a molecular weight of 8.9 kDa. The deduced amino acid sequence of the  PoCCH  gene product is 87 and 78% identical to those of tomato and  Arabidopsis , respectively, with a high degree of conservation in both the metal-binding and lysine-rich regions. However, the  PoCCH  gene product lacks the C-terminal extension identified in  Arabidopsis  CCH. Southern blot analysis suggested that the  PoCCH  gene is present in low copy numbers in poplar. The expression of  PoCCH  increased under copper deprivation conditions. The expression of  PoCCH  was down-regulated by high concentrations of copper, whereas some metals, such as aluminum and zinc, markedly induced  PoCCH , and others including cadmium, cobalt and lead had little effect on  PoCCH  expression. The plant growth regulator jasmonic acid caused an increase in  PoCCH  transcript whereas abscisic acid, salicylic acid and gibberellic acid did not. The gene was highly induced when cells were exposed to physical stress by high-speed agitation on a gyratory shaker. Other effective inducers of  PoCCH  expression in suspension culture were methyl viologen and NaCl. Thus,  PoCCH  does not respond to all stresses, but responds specifically to certain metals and abiotic stresses that induce oxidative damage. Our results suggest that JA is involved in regulating  PoCCH  expression in poplar cells.

Differential expression of a poplar copper chaperone gene in response to various abiotic stresses