Intraspecific varibility of corynespora cassiicola inferred from single nucleotide polymorphisms in ITS region of ribosomal DNA

Two single nucleotide polymorphisms (SNPs) were detected in the internal transcribed spacer region of the nuclear ribosomal DNA (rDNA-ITS) of 21 Corynespora cassiicola isolates obtained from a number of Hevea clones grown in rubber plantations in Malaysia. The two SNPs correlated with the physiological races of the isolates. Database searches yielded another 28 C. cassiicola isolates from diverse hosts and geographical regions. With this inclusion, a total of seven SNPs and two indels in the rDNA-ITS region were detected from all 49 C. cassiicola isolates. The knowledge of intraspecific variability (SNPs and indels) could prove useful in the delineation of physiological races or pathotypes of this fungus

Structural analysis of the glycoprotein allergen Hev b 4 from natural rubber latex by mass spectrometry

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

Correspondance: [email protected]. UMR DAP équipe DGBInternational audienceCorynespora 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 toxicit

Diversity of the cassiicolin gene in Corynespora cassiicola and relation with the pathogenicity in Heyea brasiliensis

Fungal Biol. ISI Document Delivery No.: 300WE Times Cited: 0 Cited Reference Count: 45 Cited References: Andrie RA, 2007, PHYTOPATHOLOGY, V97, P694, DOI 10.1094/PHYTO-97-6-0694 Atan S, 2003, J RUBBER RES, V6, P58 Barthe P, 2007, J MOL BIOL, V367, P89, DOI 10.1016/j.jmb.2006.11.086 Bendtsen JD, 2004, J MOL BIOL, V340, P783, DOI 10.1016/j.jmb.2004.05.028 Breton F., 2000, Journal of Rubber Research, V3, P115 Cai L, 2006, ANTON LEEUW INT J G, V89, P293, DOI 10.1007/s10482-005-9030-1 Carbone I, 1999, MYCOLOGIA, V91, P553, DOI 10.2307/3761358 Chee K. H., 1990, Review of Plant Pathology, V69, P423 Chee KH, 1987, P INT S PATH HEV BRA, P6 Ciuffetti LM, 2010, NEW PHYTOL, V187, P911, DOI 10.1111/j.1469-8137.2010.03362.x Collado J, 1999, NEW PHYTOL, V144, P525, DOI 10.1046/j.1469-8137.1999.00533.x Darmono TW, 1996, P WORKSH COR LEAF FA, P79 de Lamotte F, 2007, J CHROMATOGR B, V849, P357, DOI 10.1016/j.jchromb.2006.10.051 Deon M, 2012, PLANT SCI, V185, P227, DOI 10.1016/j.plantsci.2011.10.017 Deon M, 2012, FUNGAL DIVERS, V54, P87, DOI 10.1007/s13225-012-0169-6 Dixon LJ, 2009, PHYTOPATHOLOGY, V99, P1015, DOI 10.1094/PHYTO-99-9-1015 Farr DF, 2011, FUNGAL DATABASES GOLDMAN N, 1994, MOL BIOL EVOL, V11, P725 Gond SK, 2007, WORLD J MICROB BIOT, V23, P1371, DOI 10.1007/s11274-007-9375-x Halle F., 1968, ADANSONIA, V2, P475 Huelsenbeck JP, 2001, BIOINFORMATICS, V17, P754, DOI 10.1093/bioinformatics/17.8.754 Jinji P, 2007, AUST PLANT DIS NOTES, V2, P35, DOI DOI 10.1071/DN07017 Kingsland GC, 1985, ACTA HORTIC ISHS, V153, P229 Lee S, 2004, FUNGAL DIVERS, V17, P91 Liyanage AS, 1986, J RUBBER RES I SRI L, V65, P47 Newsam A, 1960, PLANT PATHOLOGY DIVI, P63 Nghia NA, 2008, MYCOPATHOLOGIA, V166, P189, DOI 10.1007/s11046-008-9138-8 Pongthep K, 1987, P IRRDB S CHIANG MAI, P1 Promputtha I, 2007, MICROB ECOL, V53, P579, DOI 10.1007/s00248-006-9117-x Qi YX, 2011, EUR J PLANT PATHOL, V130, P83, DOI 10.1007/s10658-010-9734-6 Qi YX, 2009, MOL BIOTECHNOL, V41, P145, DOI 10.1007/s12033-008-9109-9 R Development Core Team, 2009, R LANG ENV STAT COMP RAMAKRISHNAN T. S., 1961, Rubber Board Bulletin, V5, P32 Risterucci AM, 2000, THEOR APPL GENET, V101, P948, DOI 10.1007/s001220051566 Romruensukharom P., 2005, Journal of Rubber Research, V8, P38 Saha T, 2000, J RUBBER RES, V13, P1 Sallaud C, 2003, THEOR APPL GENET, V106, P1396, DOI 10.1007/s00122-002-1184-x Schoch CL, 2009, STUD MYCOL, P1, DOI 10.3114/sim.2009.64.01 Shimodaira H, 1999, MOL BIOL EVOL, V16, P1114 Silva WPK, 1998, PLANT PATHOL, V47, P267, DOI 10.1046/j.1365-3059.1998.00245.x Silva WPK, 2003, MYCOL RES, V107, P567, DOI 10.1017/S0953756203007755 Suryanarayanan TS, 2002, CAN J BOT, V80, P818, DOI [10.1139/B02-069, 10.1139/b02-069] Swofford DL, 2000, PHYLOGENETIC ANAL US Wahounou PJ, 1996, P WORKSH COR LEAF FA, P99 White TJ, 1990, AMPLIFICATION DIRECT Deon, Marine Fumanal, Boris Gimenez, Stephanie Bieysse, Daniel Oliveira, Ricardo R. Shuib, Siti Shuhada Breton, Frederic Elumalai, Sunderasan Vida, Joao B. Seguin, Marc Leroy, Thierry Roeckel-Drevet, Patricia Pujade-Renaud, Valerie IFC (Institut Francais du Caoutchouc, Paris, France); Michelin (Clermont-Ferrand, France); SIPH ('Societe Internationale de Plantations d'Heveas', Courbevoie, France); SOCFIN ('Societe Financiere des Caoutchoucs', Bruxelles, Belgium) This work was supported in part by a grant from the IFC (Institut Francais du Caoutchouc, Paris, France) and the companies Michelin (Clermont-Ferrand, France), SIPH ('Societe Internationale de Plantations d'Heveas', Courbevoie, France) and SOCFIN ('Societe Financiere des Caoutchoucs', Bruxelles, Belgium). We thank Gerald Oliver and Stephanie Massey for maintenance of rubber trees in the greenhouse. Elsevier sci ltd OxfordCorynespora cassiicola is an important plant pathogenic Ascomycete causing the damaging. Corynespora Leaf Fall (CLF) disease in rubber tree (Hama brasiliensis). A small secreted glycoprotein named cassiicolin was previously described as an important effector of C. cassiicola. In this study, the diversity of the cassiicolin-encoding gene was analysed in C. cassiicola isolates sampled from various hosts and geographical origins. A cassiicolin gene was detected in 47 % of the isolates, encoding up to six distinct protein isoforms. In three isolates, two gene variants encoding cassiicolin isoforms Cas2 and Cas6 were found in the same isolate. A phylogenetic tree based on four combined loci and elucidating the diversity of the whole collection was strongly structured by the toxin class, as defined by the cassiicolin isoform. The isolates carrying the Cas1 gene (toxin class Cas1), all grouped in the same highly supported clade, were found the most aggressive on two rubber tree cultivars. Some isolates in which no Cas gene was detected could nevertheless generate moderate symptoms, suggesting the existence of other yet uncharacterized effectors. This study provides a useful base for future studies of C. cassiicola population biology and epidemiological surveys in various host plants. (C) 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved