Differentially expressed cDNAs in Alternaria alternata treated with 2-propenyl isothiocyanate.
The molecular mechanism of the fungal tolerance phenotype to fungicides is not completely understood. This knowledge would allow for the development of environmentally friendly strategies for the control of fungal infection. With the goal of determining genes induced by 2p-ITC, a forward suppressive subtractive hybridization was performed using cDNAs from ITC-treated Alternaria alternata as a "tester" and from untreated fungus as a "driver." Using this approach, a library containing 102 ESTs was generated that resulted in 50 sequences after sequence assembly (17 contigs and 33 singletons). Blastx analysis revealed that 38% and 40% of the sequences showed significant similarity with known and hypothetical proteins, respectively, whereas 18% were not similar to known genes. These last sequences could represent novel genes that play an unknown role in the molecular responses of fungi during adaptation to 2p-ITC. Clones similar to opsins, ABC transporters, calmodulin, ATPases and SNOG proteins were identified. Using real-time RT-PCR analysis, significant inductions of an ABC transporter and a Ca(++) ATPase during 2p-ITC treatment were discovered. These results suggest that the fungal resistance phenotype to 2p-ITC involves calcium ions and 2p-ITC efflux via an ABC transporter.
Coordinación de Tecnología de Alimentos de Orígen Vegetal, Centro de Investigación en Alimentación y Desarrollo, A.C. Hermosillo, Sonora 83000, Mexico.
María Elena
ME
Báez-Flores
Rosalba
R
Troncoso-Rojas
María A Islas
MA
Osuna
Marisela Rivera
MR
Domínguez
Barry
B
Pryor
Martín Ernesto
ME
Tiznado-Hernández
D000528:Alternaria / Q000187:drug effects* / Q000235:genetics*; D018076:DNA, Complementary / Q000378:metabolism*; D025141:Drug Resistance, Fungal; D020224:Expressed Sequence Tags; D005659:Fungicides, Industrial / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D005800:Genes, Fungal; D017879:Isothiocyanates / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D018076:DNA, Complementary; D005659:Fungicides, Industrial; D017879:Isothiocyanates; C037152:isothiocyanic acid