The recombinant pea defensin Drr230a is active against impacting soybean and cotton pathogenic fungi from the genera Fusarium, Colletotrichum and Phakopsora.
Plant defensins are antifungal peptides produced by the innate immune system plants developed to circumvent fungal infection. The defensin Drr230a, originally isolated from pea, has been previously shown to be active against various entomopathogenic and phytopathogenic fungi. In the present study, the activity of a yeast-expressed recombinant Drr230a protein (rDrr230a) was tested against impacting soybean and cotton fungi. First, the gene was subcloned into the yeast expression vector pPICZαA and expressed in Pichia pastoris. Resulting rDrr230a exhibited in vitro activity against fungal growth and spore germination of Fusarium tucumaniae, which causes soybean sudden death syndrome, and against Colletotrichum gossypii var. cephalosporioides, which causes cotton ramulosis. The rDrr230a IC50corresponding to inhibition of fungal growth of F. tucumaniae and C. gossypii var. cephalosporioides was 7.67 and 0.84 µM, respectively, demonstrating moderate activity against F. tucumaniae and high potency against C. gossypii var. cephalosporioides. Additionally, rDrr230a at 25 ng/µl (3.83 µM) resulted in 100 % inhibition of spore germination of both fungi, demonstrating that rDrr230a affects fungal development since spore germination. Moreover, rDrr230a at 3 µg/µl (460.12 µM) inhibited 100 % of in vitro spore germination of the obligatory biotrophic fungus Phakopsora pachyrhizi, which causes Asian soybean rust. Interestingly, rDrr230a substantially decreased the severity of Asian rust, as demonstrated by in planta assay. To our knowledge, this is the first report of a plant defensin active against an obligatory biotrophic phytopathogenic fungus. Results revealed the potential of rDrr230a as a candidate to be used in plant genetic engineering to control relevant cotton and soybean fungal diseases.
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil.|Universidade Federal do Rio Grande do Norte, Avenida Senador Salgado Filho 3000, Lagoa Nova, Natal, RN, 59078-970, Brazil.
Ariane Ferreira
AF
Lacerda
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil.|Centro Universitário do Distrito Federal, SEP/SUL EQ 704/ A12 904 Conjunto A, Brasília, DF, 70390-045, Brazil.
Rafael Perseghini
RP
Del Sarto
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil.
Marilia Santos
MS
Silva
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil.
Erico Augusto Rosas
EAR
de Vasconcelos
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil.
Roberta Ramos
RR
Coelho
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil.|Universidade Federal do Rio Grande do Norte, Avenida Senador Salgado Filho 3000, Lagoa Nova, Natal, RN, 59078-970, Brazil.
Vanessa Olinto
VO
Dos Santos
Embrapa Soja, Rodovia Carlos João Strass, Distrito de Warta, A10 PO Box 231, Distrito de Warta, PR, 86001-970, Brazil.
Claudia Vieira
CV
Godoy
Embrapa Soja, Rodovia Carlos João Strass, Distrito de Warta, A10 PO Box 231, Distrito de Warta, PR, 86001-970, Brazil.
Claudine Dinali Santos
CDS
Seixas
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil.
Maria Cristina Mattar
MCM
da Silva
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil. fatima.grossi@embrapa.br.|Universidade Católica de Brasília, SGAN 916 Módulo B Avenida W5, Brasília, DF, 70790-160, Brazil. fatima.grossi@embrapa.br.
Maria Fatima
MF
Grossi-de-Sa
Colletotrichum gossypii var. cephalosporioides; Defensin; Fusarium tucumaniae; Phakopsora pachyrhizi; Pichia pastoris; Pisum sativum