Detection of Colletotrichum falcatum causing red rot of sugarcane by enzyme-linked immunosorbent assay

Khalid A I M Bukhari, K. Nithya, V. Valluvaparidasan, V. Paranidharan, R. Velazhahan

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Red rot disease of sugarcane caused by Colletotrichum falcatum Went is one of the most destructive diseases of sugarcane (Saccharum officinarum L.) worldwide. The pathogen spreads primarily through infected sugarcane setts and hence the use of disease-free setts is essential to prevent the disease. In order to develop immunological method for detection of C. falcatum, two proteins with molecular weights of 27 kDa and 45 kDa were purified from the mycelium of C. falcatum race Cf 05 and used as antigen source to raise polyclonal antibodies in NewZealand white rabbit. The developed polyclonal antibodies were tested for detection of C. falcatum by enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. The polyclonal antibodies specifically detected C. falcatum in extracts from infected plants, both in immunoblot and ELISA. The ELISA results showed that the developed polyclonal antibodies were highly specific to C.falcatum. The developed antibodies were very sensitive and could detect C.falcatum proteins even at a dilution of 1:50,000. Higher ELISA absorbance values were recorded even at an antigen dilution of 1:500. In western blot analysis, protein bands with molecular weights of 27 kDa and 45 kDa reacting to antisera raised against 27 kDa and 45 kDa mycelial proteins of C. falcatum, respectively, were detected in protein samples from red rot infected canes. The high specific reactivity and sensitivity of the antisera indicate its potential suitability for ELISA-based detection of C. falcatum.

Original languageEnglish
Pages (from-to)823-830
Number of pages8
JournalArchives of Phytopathology and Plant Protection
Volume45
Issue number7
DOIs
Publication statusPublished - Apr 2012

Fingerprint

Glomerella tucumanensis
sugarcane
enzyme-linked immunosorbent assay
polyclonal antibodies
proteins
antiserum
molecular weight
antigens
Saccharum officinarum
canes
absorbance
mycelium
Western blotting
rabbits
antibodies
pathogens
extracts

Keywords

  • Colletotrichum falcatum
  • ELISA
  • red rot
  • Saccharum officinarum

ASJC Scopus subject areas

  • Agronomy and Crop Science

Cite this

Detection of Colletotrichum falcatum causing red rot of sugarcane by enzyme-linked immunosorbent assay. / Bukhari, Khalid A I M; Nithya, K.; Valluvaparidasan, V.; Paranidharan, V.; Velazhahan, R.

In: Archives of Phytopathology and Plant Protection, Vol. 45, No. 7, 04.2012, p. 823-830.

Research output: Contribution to journalArticle

Bukhari, Khalid A I M ; Nithya, K. ; Valluvaparidasan, V. ; Paranidharan, V. ; Velazhahan, R. / Detection of Colletotrichum falcatum causing red rot of sugarcane by enzyme-linked immunosorbent assay. In: Archives of Phytopathology and Plant Protection. 2012 ; Vol. 45, No. 7. pp. 823-830.
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