Single-chain variable fragments antibody specific to Corynespora cassiicola toxin, cassiicolin, reduces necrotic lesion formation in Hevea brasiliensis

E. Sunderasan, Rusni A. Kadir, Valérie Pujade-Renaud, Frédéric De Lamotte, H. Y. Yeang, Sheila Nathan

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Corynespora leaf disease poses a serious threat to rubber cultivation because infected leaves develop necrotic lesions and abscise, leaving the tree unproductive. The destructiveness of Corynespora cassiicola has been largely attributed to cassiicolin, a protein toxin secreted by the fungus. Recombinant antibody technology offers hope to curtail the disease whereby single-chain variable fragments (scFv) specific to cassiicolin could bind and deactivate the toxin in genetically modified rubber trees that harbour the antibody gene. A scFv phage library was constructed from heavy and light variable chains of IgG from cassiicolin immunized Balb/C mice spleen. Biopanning of the phage library yielded a scFv clone with high specificity to cassiicolin. The nucleotide sequence and deduced amino acid sequence information of the scFv were obtained. Hemagglutinin (HA)-tagged scFv expressed in Escherichia coli is discerned as a band at ca. 30 kDa on SDS-PAGE, and the corresponding band was detected by anti-HA IgG on a Western immunoblot. Deactivation of cassiicolin by the affinity-purified scFv was demonstrated in a detached-leaf bio-assay on selected susceptible Hevea clones (PB 260, RRIM 2020, RRIM 901 and RRIM 929). The assay was also performed on clones that are relatively more resistant to the fungus (RRIM 600 and GT-1), and the results were as expected. Thus, we have successfully demonstrated that the cassicolin-specific scFv can effectively reduce cassicolin toxicity.

Original languageEnglish
Pages (from-to)19-26
Number of pages8
JournalJournal of General Plant Pathology
Volume75
Issue number1
DOIs
Publication statusPublished - Feb 2009

Fingerprint

Corynespora cassiicola
lesions (plant)
Hevea brasiliensis
toxins
DNA libraries
hemagglutinins
clones
antibodies
Corynespora
Hevea
recombinant antibodies
fungi
foliar diseases
assays
rubber
leaves
spleen
amino acid sequences
Escherichia coli
toxicity

Keywords

  • Anti-cassiicolin scFv
  • Cassiicolin
  • Corynespora cassiicola
  • Detached Hevea leaf bioassay
  • Hevea brasiliensis

ASJC Scopus subject areas

  • Plant Science
  • Agronomy and Crop Science

Cite this

Single-chain variable fragments antibody specific to Corynespora cassiicola toxin, cassiicolin, reduces necrotic lesion formation in Hevea brasiliensis. / Sunderasan, E.; Kadir, Rusni A.; Pujade-Renaud, Valérie; De Lamotte, Frédéric; Yeang, H. Y.; Nathan, Sheila.

In: Journal of General Plant Pathology, Vol. 75, No. 1, 02.2009, p. 19-26.

Research output: Contribution to journalArticle

Sunderasan, E. ; Kadir, Rusni A. ; Pujade-Renaud, Valérie ; De Lamotte, Frédéric ; Yeang, H. Y. ; Nathan, Sheila. / Single-chain variable fragments antibody specific to Corynespora cassiicola toxin, cassiicolin, reduces necrotic lesion formation in Hevea brasiliensis. In: Journal of General Plant Pathology. 2009 ; Vol. 75, No. 1. pp. 19-26.
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abstract = "Corynespora leaf disease poses a serious threat to rubber cultivation because infected leaves develop necrotic lesions and abscise, leaving the tree unproductive. The destructiveness of Corynespora cassiicola has been largely attributed to cassiicolin, a protein toxin secreted by the fungus. Recombinant antibody technology offers hope to curtail the disease whereby single-chain variable fragments (scFv) specific to cassiicolin could bind and deactivate the toxin in genetically modified rubber trees that harbour the antibody gene. A scFv phage library was constructed from heavy and light variable chains of IgG from cassiicolin immunized Balb/C mice spleen. Biopanning of the phage library yielded a scFv clone with high specificity to cassiicolin. The nucleotide sequence and deduced amino acid sequence information of the scFv were obtained. Hemagglutinin (HA)-tagged scFv expressed in Escherichia coli is discerned as a band at ca. 30 kDa on SDS-PAGE, and the corresponding band was detected by anti-HA IgG on a Western immunoblot. Deactivation of cassiicolin by the affinity-purified scFv was demonstrated in a detached-leaf bio-assay on selected susceptible Hevea clones (PB 260, RRIM 2020, RRIM 901 and RRIM 929). The assay was also performed on clones that are relatively more resistant to the fungus (RRIM 600 and GT-1), and the results were as expected. Thus, we have successfully demonstrated that the cassicolin-specific scFv can effectively reduce cassicolin toxicity.",
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