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A putative isopentenyltransferase (IPT) encoding gene was identified from a pingyitiancha ( Malus hupehensis  Rehd.) expressed sequence tag database, and the full-length gene was cloned by RACE. Based on expression profile and sequence alignment, the nucleotide sequence of the clone, named  MhIPT3 , was most similar to  AtIPT3 , an IPT gene in  Arabidopsis . The full-length cDNA contained a 963-bp open reading frame encoding a protein of 321 amino acids with a molecular mass of 37.3 kDa. Sequence analysis of genomic DNA revealed the absence of introns in the frame. Quantitative real-time PCR analysis demonstrated that the gene was expressed in roots, stems and leaves. Application of nitrate to roots of nitrogen-deprived seedlings strongly induced expression of  MhIPT3  and was accompanied by the accumulation of cytokinins, whereas  MhIPT3  expression was little affected by ammonium application to roots of nitrogen-deprived seedlings. Application of nitrate to leaves also up-regulated the expression of  MhIPT3  and corresponded closely with the accumulation of isopentyladenine and isopentyladenosine in leaves.

tree physiology

Molecular cloning of a putative gene encoding isopentenyltransferase from pingyitiancha ( Malus hupehensis ) and characterization of its response to nitrate