|DURESSA, DECHASSA - Former ARS Employee|
|CHEN, DONGQUAN - University Of Alabama|
|KLIMES, ANNA - Western New England University|
|GARCIA-PEDRAJAS, MARIA - Spanish National Research Council|
|DOBINSON, KATHERINE - Agriculture And Agri-Food Canada|
Submitted to: Verticillium International Symposium Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 3/10/2013
Publication Date: 5/6/2013
Citation: Klosterman, S.J., Duressa, D., Anchieta, A.G., Chen, D., Klimes, A., Garcia-Pedrajas, M.D., Dobinson, K.F. 2013. High throughput analysis of gene expression in microsclerotia of Verticillium dahliae. In: Koopmann, B., von Tiedemann, A., editors. Proceedings of the 11th International Verticillium Symposium. German Phytomedical Society, DPG-Verlag. p. 32.
Technical Abstract: The processes of microsclerotia formation, maintenance, and germination are critically important in the disease cycle of V. dahliae. To shed additional light on the molecular processes involved in microsclerotia biogenesis and melanin synthesis in V. dahliae, three replicate RNA-seq libraries were prepared from 10 day-old MS-producing cultures of V. dahliae (ave = 52.2 million reads), and those not producing microsclerotia (NoMS, ave = 50.6 million reads) and analyzed for differential gene expression. The comparisons revealed up-regulation of MS library genes involved in melanogenesis, including tetrahydroxynaphthalene reductase (344-fold increase) and scytalone dehydratase (231-fold increase), and additional genes located in a 48.8 kilobase melanin biosynthetic cluster. Numerous hypothetical protein-encoding genes were also identified as differentially expressed in the MS library. Differential expression of selected genes identified as up- or down-regulated by RNA-seq were analyzed by RT-qPCR from several MS and NoMs culture types, including MS cultures that were stored for 6 months, and a 7 day culture having an intermediate level of fully melanized MS. These data provide further insight into gene expression during melanin biosynthesis and MS formation in V. dahliae and may suggest targets for alternative disease control methods.