Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides

Siela N. Maximova, Jean Philippe Marelli, Ann Young, Sharon Pishak, Joseph A. Verica, Mark J. Guiltinan

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Theobroma cacao L. plants over-expressing a cacao class I chitinase gene (TcChi1) under the control of a modified CaMV-35S promoter were obtained by Agrobacterium-mediated transformation of somatic embryo cotyledons. Southern blot analysis confirmed insertion of the transgene in eight independent lines. High levels of TcChi1 transgene expression in the transgenic lines were confirmed by northern blot analysis. Chitinase activity levels were measured using an in vitro fluorometric assay. The transgene was expressed at varying levels in the different transgenic lines with up to a sixfold increase of endochitinase activity compared to non-transgenic and transgenic control plants. The in vivo antifungal activity of the transgene against the foliar pathogen Colletotrichum gloeosporioides was evaluated using a cacao leaf disk bioassay. The assay demonstrated that the TcChi1 transgenic cacao leaves significantly inhibited the growth of the fungus and the development of leaf necrosis compared to controls when leaves were wound inoculated with 5,000 spores. These results demonstrate for the first time the utility of the cacao transformation system as a tool for gene functional analysis and the potential utility of the cacao chitinase gene for increasing fungal pathogen resistance in cacao.

Original languageEnglish (US)
Pages (from-to)740-749
Number of pages10
JournalPlanta
Volume224
Issue number4
DOIs
StatePublished - Sep 1 2006

Fingerprint

Colletotrichum
MHC Class I Genes
Theobroma cacao
Cacao
Glomerella cingulata
chitinase
transgenes
pathogens
Transgenes
Chitinases
genetically modified organisms
genes
leaves
leaf development
plant damage
assays
somatic embryos
Agrobacterium
Northern blotting
Southern blotting

All Science Journal Classification (ASJC) codes

  • Genetics
  • Plant Science

Cite this

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title = "Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides",
abstract = "Theobroma cacao L. plants over-expressing a cacao class I chitinase gene (TcChi1) under the control of a modified CaMV-35S promoter were obtained by Agrobacterium-mediated transformation of somatic embryo cotyledons. Southern blot analysis confirmed insertion of the transgene in eight independent lines. High levels of TcChi1 transgene expression in the transgenic lines were confirmed by northern blot analysis. Chitinase activity levels were measured using an in vitro fluorometric assay. The transgene was expressed at varying levels in the different transgenic lines with up to a sixfold increase of endochitinase activity compared to non-transgenic and transgenic control plants. The in vivo antifungal activity of the transgene against the foliar pathogen Colletotrichum gloeosporioides was evaluated using a cacao leaf disk bioassay. The assay demonstrated that the TcChi1 transgenic cacao leaves significantly inhibited the growth of the fungus and the development of leaf necrosis compared to controls when leaves were wound inoculated with 5,000 spores. These results demonstrate for the first time the utility of the cacao transformation system as a tool for gene functional analysis and the potential utility of the cacao chitinase gene for increasing fungal pathogen resistance in cacao.",
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Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides. / Maximova, Siela N.; Marelli, Jean Philippe; Young, Ann; Pishak, Sharon; Verica, Joseph A.; Guiltinan, Mark J.

In: Planta, Vol. 224, No. 4, 01.09.2006, p. 740-749.

Research output: Contribution to journalArticle

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T1 - Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides

AU - Maximova, Siela N.

AU - Marelli, Jean Philippe

AU - Young, Ann

AU - Pishak, Sharon

AU - Verica, Joseph A.

AU - Guiltinan, Mark J.

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