02635naa a2200385 a 450000100080000000500110000800800410001902200140006002400390007410000200011324501310013326000090026450001170027352014100039065000190180065000300181965000100184965000220185965000100188165000200189165300130191165300320192465300260195665300220198265300210200470000230202570000220204870000160207070000150208670000230210170000170212470000220214170000260216377300600218918652532017-05-30       2010    bl uuuu     u00u1 u    #d  a0032-09357 aDOI 10.1007/s00425-010-1272-02DOI1 aPIROVANI, C. P.  aTheobroma cacao cystatins impair Moniliophthora perniciosa mycelial growth and are involved in postponing cell death symptoms.  c2010  aDisponível em: &lthttp://www.springerlink.com/content/c41483451417h040/fulltext.pdf&gt. Acesso em 26 out. 2010.  aThree cystatin open reading frames named TcCys1, TcCys2 and TcCys3 were identified in cDNA libraries from compatible interactions between Theobroma cacao (cacao) and Moniliophthora perniciosa. In addition, an ORF named TcCys4 was identified in the cDNA library of the incompatible interaction. The cDNAs encoded conceptual proteins with 209, 127, 124, and 205 amino acid residues, with a deduced molecular weight of 24.3, 14.1, 14.3 and 22.8 kDa, respectively. His-tagged recombinant proteins were purified from Escherichia coli expression, and showed inhibitory activities against M. perniciosa. The four recombinant cystatins exhibited Ki values against papain in the range of 152–221 nM. Recombinant TcCYS3 and TcCYS4 immobilized in CNBr–Sepharose were efficient to capture M. perniciosa proteases from culture media. Polyclonal antibodies raised against the recombinant TcCYS4 detected that the endogenous protein was more abundant in young cacao tissues, when compared with mature tissues. A *85 kDa cacao multicystatin induced by M. perniciosa inoculation, MpNEP (necrosis and ethylene-inducing protein) and M. perniciosa culture supernatant infiltration were detected by anti-TcCYS4 antibodies in cacao young tissues. A direct role of the cacao cystatins in the defense against this phytopathogen was proposed, as well as its involvement in the development of symptoms of programmed cell death.  aMoniliophthora  aMoniliophthora perniciosa  aCacau  aDoença de Planta  aFungo  aTheobroma Cacao  aCystatin  aCysteine protease inhibitor  aProgrammed cell death  aVassoura-de-bruxa  aWitches’ broom1 aSANTIAGO, A. da S.1 aSANTOS, L. S. dos1 aMICHELI, F.1 aMARGIS, R.1 aGESTEIRA, A. da S.1 aALVIM, F. C.1 aPEREIRA, G. A. G.1 aCASCARDO, J. C. de M.  tPlanta, Heidelberggv. 232, n. 6, p. 1485–1497, 2010.