Two new cysteine proteinases with specific expression patterns in mature and senescent tobacco (Nicotiana tabacum L.) leaves

Cysteine proteinases are involved in various physiological and developmental processes in plants. Two cDNAs from senescent and non-senescent tobacco leaves were isolated with degenerate primers designed from conserved regions of plant senescence-associated cysteine proteinases using rapid amplification of cDNA ends (RACE). Both sequences encode papain-like cysteine proteinases: the 833 bp fragment (NtCP1) encoding a C-terminus partial sequence of a putative tobacco cysteine proteinase gene whereas the 1300 bp fragment (NtCP2) is a full-length cysteine proteinase. On the amino acid sequence level, NtCP1 has a high similarity with other senescence-associated cysteine proteinases. It is expressed only in senescent leaves. It is not induced in mature green leaves upon exposure to drought or heat. These results suggest that it might be a good developmental senescence marker in tobacco. By contrast, NtCP2 has a high similarity to KDEL-tailed cysteine proteinases and is expressed in mature green leaves. Both drought and heat decreased NtCP2 transcript abundance in mature green leaves. It is concluded that NtCP1 is a senescence-specific cysteine proteinase whereas NtCP2 fulfils roles in green leaves that might be similar to those of KDEL-tailed cysteine proteinases involved, for example, in programmed cell death.