PMID- 36867585 OWN - NLM STAT- Publisher LR - 20240216 IS - 0191-2917 (Print) IS - 0191-2917 (Linking) DP - 2023 Mar 3 TI - First identification of Xanthomonas nasturtii as the cause of black rot of watercress in Hawaii. LID - 10.1094/PDIS-05-22-1043-PDN [doi] AB - Watercress (Nasturtium officinale) has been in continuous production in Hawaii for over a century and is part of the local diet. Black rot of watercress was first identified as caused by Xanthomonas nasturtii in Florida (Vicente et al., 2017), but symptoms of this disease have also been regularly observed in Hawaii production in all islands, mostly during the rainy season from December to April in areas with poor air circulation (McHugh & Constantinides, 2004). Initially, this disease was attributed to X. campestris due to similar symptoms to black rot of brassicas. Samples of watercress with symptoms that could be attributed to a bacterial disease including yellow spots and lesions on leaves and stunting and deformation of plants in more advanced stages, were collected from a farm in Aiea in the island of Oahu, Hawaii, in October 2017. Isolations were performed at the University of Warwick. Fluid from macerated leaves was streaked into plates of King's B (KB) medium and Yeast Dextrose Calcium Carbonate Agar (YDC). After 48-72 hrs incubation at 28 degrees C, the plates showed a range of mixed colonies. Single cream-yellow mucoid colonies were sub-cultured several times and pure isolates including WHRI 8984 were stored at -76 degrees C as previously described (Vicente et al., 2017). Colony morphology was observed in KB plates and, in contrast to the type strain from Florida (WHRI 8853 = NCPPB 4600), isolate WHRI 8984 did not cause browning of the medium. Pathogenicity was tested on four-week old watercress and Savoy cabbage cv. Wirosa F1 plants by inoculations on leaves as previously described (Vicente et al., 2017). WHRI 8984 did not produce symptoms when inoculated on cabbage but produced typical symptoms on watercress. A re-isolation from a leaf showing a V-shaped lesion, produced isolates with the same morphology, including isolate WHRI 10007A, that was also shown to be pathogenic to watercress therefore completing the Koch's postulates. Fatty acid profiling was performed on WHRI 8984 and 10007A and controls grown on trypticase soy broth agar (TSBA) plates at 28 degrees C for 48 hrs as described by Weller et al. (2000). Profiles were compared with the RTSBA6 v6.21 library; as the database does not include X. nasturtii, the results were only interpreted at the genus level, and both isolates were shown to be Xanthomonas sp. For molecular analysis, DNA was extracted and the gyrB partial gene was amplified and sequenced as described by Parkinson et al. (2007). Comparisons with sequences available in the National Centre for Biotechnology Information (NCBI) databases using the Basic Local Alignment Search Tool (BLAST) showed that partial gyrB of WHRI 8984 and 10007A were identical to the type strain from Florida therefore confirming that they belong to X. nasturtii. For whole genome sequencing, genomic libraries for WHRI 8984 were prepared using Illumina's Nextera XT v2 kit and sequenced on a HiSeq Rapid Run flowcell. The sequences were processed as previously described (Vicente et al., 2017) and the whole genome assembly has been deposited in GenBank (accession QUZM00000000.1); the phylogenetic tree shows that WHRI 8984 is close, but not identical to the type strain. This is the first identification of X. nasturtii in watercress crops in Hawaii. Control of this disease generally involves the use of copper bactericides and minimizing moisture on leaves by reducing overhead irrigation and increasing air circulation (McHugh & Constantinides, 2004); seed testing might help to select batches that are disease free and, in longer term, breeding for disease resistance might produce cultivars that can be part of management strategies. FAU - Vicente, Joana G AU - Vicente JG AD - Fera Science Ltd, 16233, York, United Kingdom of Great Britain and Northern Ireland. AD - University of Warwick, 2707, School of Life Sciences, Coventry, United Kingdom of Great Britain and Northern Ireland; joana.vicente@fera.co.uk. FAU - McHugh, John AU - McHugh J AD - Plant Industry Division, Department of Agriculture, Honolulu, Hawaii, United States; jjmchughjr@gmail.com. FAU - Bryning, Adam AU - Bryning A AD - Fera Science Ltd, 16233, York, United Kingdom of Great Britain and Northern Ireland; adam.bryning@fera.co.uk. FAU - Carroll, Sarah AU - Carroll S AD - Fera Science Ltd, 16233, York, United Kingdom of Great Britain and Northern Ireland; sarah.carroll@fera.co.uk. FAU - Harrison, James AU - Harrison J AD - University of Exeter, 3286, College of Life and Environmental Sciences, Exeter, United Kingdom of Great Britain and Northern Ireland; J.W.Harrison@exeter.ac.uk. FAU - Studholme, David AU - Studholme D AD - University of Exeter, 3286, College of Life and Environmental Sciences, Exeter, United Kingdom of Great Britain and Northern Ireland; d.j.studholme@exeter.ac.uk. LA - eng PT - Journal Article DEP - 20230303 PL - United States TA - Plant Dis JT - Plant disease JID - 9882809 SB - IM OTO - NOTNLM OT - Nasturtium officinale OT - Xanthomonas nasturtii OT - Fatty Acid Profiling OT - black rot OT - gyrB OT - identification OT - sequencing OT - watercress EDAT- 2023/03/04 06:00 MHDA- 2023/03/04 06:00 CRDT- 2023/03/03 13:13 PHST- 2023/03/03 13:13 [entrez] PHST- 2023/03/04 06:00 [pubmed] PHST- 2023/03/04 06:00 [medline] AID - 10.1094/PDIS-05-22-1043-PDN [doi] PST - aheadofprint SO - Plant Dis. 2023 Mar 3. doi: 10.1094/PDIS-05-22-1043-PDN.