Identification of genes associated with asexual reproduction in Phyllosticta citricarpa mutants obtained through Agrobacterium tumefaciens transformation.
Phyllosticta citricarpa is the epidemiological agent of Citrus Black Spot (CBS) disease, which is responsible for large economic losses worldwide. CBS is characterized by the presence of spores (pycnidiospores) in dark lesions of fruit, which are also responsible for short distance dispersal of the disease. The identification of genes involved in asexual reproduction of P. citricarpa can be an alternative for directional disease control. We analyzed a library of mutants obtained through Agrobacterium tumefaciens transformation system, looking for alterations in growth and reproductive structure formation. Two mutant strains were found to have lost the ability to form pycnidia. The flanking T-DNA insertion regions were identified on P. citricarpa genome by using blast analysis and further gene prediction. The predicted genes containing the T-DNA insertions were identified as Spindle Poison Sensitivity Scp3, Ion Transport protein, and Cullin Binding proteins. The Ion Transport and Cullin Binding proteins are known to be correlated with sexual and asexual reproduction in fungi; however, the exact mechanism by which these proteins act on spore formation in P. citricarpa needs to be better characterized. The Scp3 proteins are suggested here for the first time as being associated with asexual reproduction in fungus. This protein is associated with microtubule formation, and as microtubules play an essential role as spindle machinery for chromosome segregation and cytokinesis, insertions in this gene can lead to abnormal formations, such as that observed here in P. citricarpa. We suggest these genes as new targets for fungicide development and CBS disease control, by iRNA.
Department of Genetics, Universidade Federal do Paraná, P.O. BOX 19071, CEP: 81531-980 Curitiba, PR, Brazil.
Eduardo Henrique
EH
Goulin
Department of Genetics, Universidade Federal do Paraná, P.O. BOX 19071, CEP: 81531-980 Curitiba, PR, Brazil.
Daiani Cristina
DC
Savi
Department of Genetics, Universidade Federal do Paraná, P.O. BOX 19071, CEP: 81531-980 Curitiba, PR, Brazil.
Desirrê Alexia Lourenço
DAL
Petters
Department of Genetics, Universidade Federal do Paraná, P.O. BOX 19071, CEP: 81531-980 Curitiba, PR, Brazil.
Vanessa
V
Kava
Department of Genetics, Universidade Federal do Paraná, P.O. BOX 19071, CEP: 81531-980 Curitiba, PR, Brazil.
Lygia
L
Galli-Terasawa
Fund for Citrus Protection, Fundecitrus, Av. Dr. Adhemar Pereira de Barros, 201, CEP: 14807-040 Araraquara, SP, Brazil.
Geraldo José
GJ
Silva
Department of Genetics, Universidade Federal do Paraná, P.O. BOX 19071, CEP: 81531-980 Curitiba, PR, Brazil. Electronic address: ch.glienke@gmail.com.
Chirlei
C
Glienke
D016960:Agrobacterium tumefaciens / Q000502:physiology*; D001203:Ascomycota / Q000502:physiology*; D002957:Citrus / Q000382:microbiology; D004269:DNA, Bacterial; D015966:Gene Expression Regulation, Fungal; D023061:Gene Order; D017930:Genes, Reporter; D056265:Microbial Interactions; D016254:Mutagenesis, Insertional; D009154:Mutation; D010935:Plant Diseases / Q000382:microbiology; D012100:Reproduction, Asexual / Q000235:genetics*; D014170:Transformation, Genetic
D004269:DNA, Bacterial; C036483:T-DNA
Agrobacterium tumefaciens; Asexual reproduction; Cullin binding protein; Ion transport protein; Phyllosticta citricarpa; Spindle poison sensitivity Spc3