African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12269

Full Length Research Paper

Recovery of active pathogenesis-related enzymes from the apoplast of Musa acuminata infected by Mycosphaerella fijiensis

Ignacio Islas-Flores*
  • Ignacio Islas-Flores*
  • Unidad de Bioquimica y Biologia Molecular de Plantas, Calle 43 No. 130, Colonia Chuburna de Hidalgo, 97200 Merida, Yucatan, Mexico.
  • Google Scholar
Carmela Alcocer-Alvarez
  • Carmela Alcocer-Alvarez
  • Unidad de Bioquimica y Biologia Molecular de Plantas, Calle 43 No. 130, Colonia Chuburna de Hidalgo, 97200 Merida, Yucatan, Mexico.
  • Google Scholar
Yasmín Abril Sánchez-Rodriguez
  • Yasmín Abril Sánchez-Rodriguez
  • Unidad de Bioquimica y Biologia Molecular de Plantas, Calle 43 No. 130, Colonia Chuburna de Hidalgo, 97200 Merida, Yucatan, Mexico.
  • Google Scholar
Blondy Canto-Canche
  • Blondy Canto-Canche
  • Unidad de Biotecnologia, Centro de Investigacion Cientifica de Yucatan A.C., Calle 43 No. 130, Colonia Chuburna de Hidalgo, 97200 Merida, Yucatan, Mexico.
  • Google Scholar


  •  Received: 25 November 2014
  •  Accepted: 11 May 2015
  •  Published: 10 June 2015

Abstract

The fungus Mycosphaerella fijiensis causes black Sigatoka (BS) disease, a major pathogen in the banana industry worldwide. Numerous molecular and biochemical studies have been done for the M. fijiensis, Musa acuminata interaction, but this is the first study describing the zymographic behavior of β-1,3-glucanase, chitinase and protease in the apoplast and symplast of healthy, BS-infected but asymptomatic and BS-diseased banana leaves. In BS-infected tissues, β-1,3-glucanase enzymatic activity was associated with two polypeptides with retention index (Ri) values of 0.43 and 0.56. These were more notable in the apoplast than in the symplast. Chitinase activity in BS-infected tissue in both the apoplast and symplast was mainly associated with a single polypeptide (Ri = 0.89). Both β-1,3-glucanase and chitinase activities were apparently more intense in BS-infected leaves than in healthy leaves. Protease activity was associated with two polypeptides (Ri = 0.04 and 0.14). In both the apoplast and symplast, the Ri 0.04 polypeptide increased in intensity with disease progression, whereas Ri 0.14 polypeptide intensity decreased. Overall protease activity intensity was higher in the symplast. Maximum symplast contamination of the apoplast was 2% as estimated by glucose 6-phosphate dehydrogenase activity, a biochemical marker for symplast. Accumulation of pathogenesis-related enzymatic activities in the apoplast of M. acuminata leaf tissue was caused by host-controlled enzyme downloading in response to M. fijiensis infection. Clear differences were identified in the electrophoretic profiles of healthy and diseased banana plants. The results further support a putative role of these enzymes in the extracellular defense repertoire of banana and, more importantly, suggest that M. fijiensis possesses a mechanism for suppression and delay of defense response in M. acuminata.

 

Key words: Black Sigatoka, glucose 6-phosphate dehydrogenase, pathogenesis-related (PR) proteins, polyacrylamide gel electrophoresis (PAGE), retention index (Ri), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Abbreviation

BS, Black Sigatoka; PAGE, polyacrylamide gel electrophoresis; Ri, retention index; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; PCR, polymerase chain reaction; CTAB, cetyltrimethyl ammonium bromide; PVPP, polyvinyl polypyrrolidone; G6PD, glucose 6-phosphate dehydrogenase.