The AjTOX2 genes have been deposited in GenBank with accession numbers KC862269-KC862275 (Additional file 1: Table S1). Virulence assays Virulence assays on maize, cabbage, Arabidopsis thaliana, and Fumana procumbens were performed with spores collected from V8-juice plates with 0.1% Tween-20.
The spore concentration was adjusted to ~105 spores/ml. For maize, six- week old plants (genotype hm1/hm1 or HM1/HM1) were spray-inoculated and the plants covered with plastic bags overnight to maintain humidity, after which the plants were grown in a greenhouse. Observations of disease progression BIBF 1120 were made beginning 3 d post-inoculation. For cabbage (Brassica oleracea), plants were grown in a growth chamber at 20°C, 70% relative humidity, and a 12-hr light /dark cycle. Leaves from 4-week-old plants were spot-inoculated with 10 μl of inoculum. Plants were covered overnight find more to maintain humidity. Plants were observed for signs of infection beginning 4 d after inoculation. For Arabidopsis, plants (Col-0, a pad3 near-isogenic mutant, and a DELLA quadruple mutant ) were grown in a growth chamber at 20°C, 70% relative humidity, and a 12-hr light/dark cycle. The third through the seventh true leaves from 4-week-old
plants were spot-inoculated with 10 μl of spores. Plants were covered overnight to maintain humidity and observed for signs of infection starting 4 d after inoculation. Seeds of Fumana procumbens were obtained from Hardyplants, Apple Valley, MN, and after scarification with a razor blade were germinated in glass scintillation vials on Whatman #1 filter paper. Seven to ten day-old seedlings were triclocarban transferred to soil and grown at room temperature under a 32 watt fluorescent light (Philips 432T8/TL741 Universal/ Hi-Vision Hg). Conidial suspensions of A. jesenskae (10 μl) were applied as a drop on the surface of leaves of 5-6 month old plants. Plants were covered with a clear plastic dome lid and kept at 100% relative humidity for 48 hr. Observations were made beginning 3 d after inoculation. Acknowledgements This work was supported by award DE-FG02-91ER20021 from
the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, Office of Science, U.S. Department of Energy. We thank Dr. Emory Simmons (Wabash College, Crawfordsville, Indiana) for the strain of A. jesenskae. We thank Dr. Gerald Adams (University of Nebraska) for advice on growing A. jesenskae, the MSU Research Technology Support Facility for the DNA sequencing, and the MSU Mass Spectrometry Core Facility for the mass spectrometry. Electronic supplementary material Additional file 1: Conservation of the genes for HC-toxin https://www.selleckchem.com/products/LDE225(NVP-LDE225).html biosynthesis in Alternaria jesenskae . Table S1. GenBank accession numbers for genes of TOX2 and AjTOX2. Table S2. List of primers used to amplify probes used for Southern blots (Figure 2). (DOCX 14 KB) References 1. Walton JD: Host-selective toxins: agents of compatibility.