Abstract
Abstract: Cotton Ramulosis (Gossypium hirsutum) is an important disease affecting cotton plantations in Brazil, and its causal agent, Colletotrichum gossypiivar.cephalosporioides(Cgc), according to the Brazilian phytosanitary authority, was considered a regulated non quarantine pest. It makes this microorganism subject to standardization in seed certification programs. The current seed health testing for detecting that pathogen in seed samples does not provide reliable results for routine analysis. On this paper, attempts were made to design specific primers for detection of Cgc associated with cotton seed. Two primer sets were selected based on the analysis of a multiple alignment of gene’s sequence encoding the glyceraldehyde 3-phosphate dehydrogenase from Cgc, C. gossypii and reference strains of the C. gloeosporioides species complex. The conserved sites unique to Cgc strains were used to design specific fragment of 140 bp. The primer specificity was confirmed by using other fungi. The primers produced a detectable band of target DNA of Cgc in all inoculum potentials of the pathogen artificially inoculated by the water restriction technique. The developed primer pair represents, therefore, a reliable and rapid mean to diagnose the Ramulosis agent in cotton seed.
Highlights
Ramulosis is one of the most prominent diseases in cotton (Gossypium hirsutum) in Brazil, and it is caused by Colletotrichum gossypii var. cephalosporioides A
Isolates obtention: Colletotrichum gossypii var. cephalosporioides isolates and other fungi species were obtained from the mycological collection of the Mycology Laboratory and of the Seed Pathology Laboratory of the Universidade Federal de Lavras (UFLA), in Lavras, MG, Brazil (Table 1)
Colletotrichum gossypii var. cephalosporioides specific primers designed from the GAPDH gene had the following sequences: CGC1F (5’- CAG ACT ACA AGG CCA ACG C- 3’) and CGC1R (5’- GAG TCG TAC TTG AGC ATG TAG- 3’)
Summary
Ramulosis is one of the most prominent diseases in cotton (Gossypium hirsutum) in Brazil, and it is caused by Colletotrichum gossypii var. cephalosporioides A. This organism belongs to the Colletotrichum gloeosporioides species complex, as well as Colletotrichum gossypii South. (Cg), which causes Anthracnose in cotton (Salustiano et al, 2014) Cephalosporioides and C. gossypii) belong to the Ascomycota phylum, having as main feature the production of conidial mass with orange color in acervuli and conidia morphologically similar (Bailey et al, 1996). Both pathogens are transmitted by seeds and cause damages in cotton plants (Silva-Mann et al, 2005; Mehta and Mehta, 2010)
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