First Report of Xanthomonas hortorum pv. carotae Causing Bacterial Leaf Blight of Carrot in Mauritius.

Bacterial blight of carrot (Daucus carota) is caused by Xanthomonas hortorum pv. carotae (4). The pathogen is seed transmitted and carrot seeds can be an important source of primary inoculum (2). A 2008-2009 outbreak of a disease resembling bacterial blight was observed in Mauritius in 10 ha of carrot crops, primarily in humid areas of the island, at an estimated incidence of 10%. Carrot leaves with angular, water-soaked leaf spots that turned necrotic were collected at Plaine Sophie, Mauritius in December 2008. Yellow, Xanthomonas-like colonies were isolated onto KC agar medium (3). MultiLocus sequence analysis (MLSA) with four genes (atpD, dnaK, efp, and gyrB) was performed as described previously (1) on five carrot strains together with two reference strains of X. hortorum pv. carotae (LMG 8643 and LMG 8644). The reference strains were identical. Of the five Mauritius strains, two (LG1-1 and LG1-4) were identical, and most closely related to, but distinct from, the reference strains (genetic distance of 0.02). The other three strains represented two sequence types identified as Xanthomonas sp. based on a phylogenetic tree derived from concatenated sequences, but were not related to any type strain. PCR assays with a 3S primer pair specific for X. hortorum pv. carotae (2) produced an amplicon of approximately 350 bp from isolates LG1-1, LG1-4, and each of the reference strains. A PCR assay with a 9B primer pair (2) yielded an amplicon of 0.9 kb for strains LG1-1, LG1-4, and LMG 8644, whereas LMG 8643 yielded an amplicon of approximately 2.0 kb (2). Foliage of 4-week-old plants (36 plants per strain) of the carrot cv. Senator F1 were spray inoculated with a suspension of each strain using an 18-h culture in sterile 0.01 M tris buffer (pH 7.2) with approximately 1 × 108 CFU/ml. Plants sprayed with tris buffer were used as a negative control treatment. Plants were incubated in a growth chamber at 26 ± 1°C at a relative humidity of 95 ± 5% and a photoperiod of 16 h. Water-soaked lesions that developed into necrotic areas were observed 12 to 15 days after inoculation of LG1-1, LG1-4, and the two reference strains. Bacteria were recovered from lesions onto KC medium (3) 3 weeks after inoculation with mean Xanthomonas populations of at least 1 × 107 CFU/lesion. Colonies with morphology typical of Xanthomonas were recovered and typed using atpD sequencing to fulfill Koch's postulates. Although Xanthomonas-like bacteria were isolated from symptomatic carrot leaves in Mauritius in 1989, the results of that study were not published. To our knowledge, this is the first report of molecular and pathological characterization of this pathogen in carrot crops in Mauritius. References: (1) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (2) X. Q. Meng et al. Plant Dis. 88:1226, 2004. (3) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (4) L. Vauterin et al. Int. J. Syst. Bacteriol. 45:472, 1995.

First Report of Iris yellow spot virus in Onion in Mauritius.

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) transmitted by thrips (Thrips tabaci Lindeman) is an economically important viral pathogen of bulb and seed onion (Allium cepa) crops in many onion-growing areas of the world (2,3). In Africa, IYSV has been reported in Reunion (4) and South Africa (1). In June 2008, diamond-shaped lesions that are typical of IYSV were observed on onion seed scapes in an onion plot of 0.25 ha at Reduit in the central part of Mauritius. Disease incidence was 80% with a severity of 50 to 75% of the scape surface area. Lodging was observed in 25% of the symptomatic plants. Twenty-two symptomatic plants were tested and found to be positive for IYSV when tested by double antibody sandwich (DAS)-ELISA with a commercially available kit (Agdia Inc., Elkhart, IN). The presence of the virus was confirmed by reverse transcription (RT)-PCR tests with primers 917L: 5'-TAAAACTTAACTAACACAAA-3' and 56U: 5'-TCCTAAGTATTCACCAT-3' as forward and reverse primers, respectively, for specific sequences flanking the CP gene. Another set of primers specific to the small (S) RNA of IYSV (5'-TAAAACAAACATTCAAACAA-3' and 5'-CTCTTAAACACATTT AACAAGCAC-3') produced an amplicon of approximately 1.2 kb that includes the 772-bp nucleocapsid (N) gene. The 1.2-kb amplicon was cloned and four clones were sequenced and consensus sequence was used for comparisons. Sequence analysis showed that the N gene of the IYSV isolate from Mauritius (GenBank Accession No. HM218822) shared the highest nucleotide sequence identity (99%) with several known IYSV N gene sequences (Accession Nos. FJ785835 and AM900393) available in the GenBank, confirming the presence of IYSV in the onion crops in Mauritius. A survey was subsequently carried out from July to November 2008 in major onion-growing localities at La Marie, Henrietta, Reduit, and Plaine Sophie (center); Bassin, La Ferme, and La Chaumiere (west); Grand Sable, Petit Sable, and Plaisance (south, southeast); and Belle Mare, Trou d'Eau Douce, and Palmar (east) to monitor the distribution of the disease on the island. Symptomatic samples with diamond-to-irregularly shaped lesions were observed and 155 symptomatic and 35 nonsymptomatic samples were collected and screened by DAS-ELISA for IYSV and Tomato spotted wilt virus (TSWV), another tospovirus reported to infect onion elsewhere. Sixty-six percent of the symptomatic samples screened (102 of 155) tested positive for IYSV. No IYSV was detected in the symptomless samples. There was no serological indication of TSWV infection in the samples. Samples that tested positive for IYSV were collected from Belle mare, Palmar, and Trou d'eau douce in the east and La Ferme in the west. Cultivars infected were Gandiole, Local Red, and Veronique. No IYSV was detected in the bulbs. The vector, T. tabaci, was observed in infected onion parcels surveyed and is known to occur in all onion-producing areas of the island. To our knowledge, this is the first report of IYSV in onion in Mauritius. Further surveys and monitoring of IYSV incidence, along with its impact on the yield, need to be established. References: (1) L. J. du Toit et al. Plant Dis. 91:1203, 2007. (2) D. H. Gent et al. Plant Dis. 88:446, 2004. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) I. Robène-Soustrade et al. Plant Pathol. 55:288, 2006.

First Report of Tomato chlorosis virus in Tomato on Mauritius Island.

In February of 2007, a virus disease survey on tomato plants (Solanum lycopersicum) in greenhouses and open fields was conducted on the island of Mauritius at the request of the Agricultural Research and Extension Unit (AREU), sponsored by the European Union, and funded by the Programme Régional de Protection des Végétaux (PRPV). Yellowing symptoms on the lower and middle leaves of tomato plants and whiteflies (Bemisia tabaci) were observed in greenhouses in Pailles, located in the north region of the island. The interveinal chlorosis pattern of the discolored leaves was similar to symptoms described for Tomato chlorosis virus (ToCV; genus Crinivirus) detected on tomato in 2004 on Reunion Island (1), suggesting the possible involvement of the same virus. Six symptomatic tomato leaf samples were collected from separate plants in the Pailles greenhouses. Total RNA was extracted from these samples with the Qiagen (Courtaboeuf, France) RNeasy Plant Mini Kit. Reverse transcription-PCR was used for molecular diagnosis, independently using two sets of specific ToCV primers. The first set of primers, ToCV-172 and ToCV-610, was designed to amplify the highly conserved region of the heat shock protein 70 (HSP70) gene (2). The second set of primers was designed to amplify the coat protein (CP) gene (forward-CP-ToCV-4384: 5'-ATCCTCTGGTTAGACCGTTAG-3' and reverse as in Segev et al. [3]). PCR products of the expected size (439 and 725 bp, respectively) were observed for the six samples from the greenhouse from Pailles. For each set of primers, two PCR products obtained from two different samples were cloned using the pGEM-T Easy Vector system (Promega, Madison, WI) and sequenced (Macrogen, Seoul, Korea). The two HSP70 sequences (GenBank-EMBL-DDBJ Accession Nos. AM884013 and AM884014) and the two CP sequences (FM206381 and FM206382) had 100% nucleotide identities (DNAMAN; Lynnon BioSoft, Quebec City, Canada). The highest nucleotide identities of the 439-bp fragment of HSP70 gene (NCBI, BLASTn) were 97% with ToCV isolates from France (DQ355214, DQ355215, and DQ355216), Florida (AY903448), Italy (AM231038 and AY048854), Mayotte Island (AM748818), Portugal (AF234029), and Reunion Island (AM748816). Similarly, the highest nucleotide identities (98%) were obtained with ToCV isolates from France (EU625350) and Spain (DQ136146), with the 725-bp fragments of CP gene. Interestingly, ToCV isolates from Mauritius and Reunion are as divergent as isolates from the rest of the world, which suggests the possibility of different introductions. In conclusion, observed symptoms and laboratory results based on two different regions of the genome confirm the presence of ToCV in symptomatic tomatoes on the island of Mauritius, for the first time to our knowledge. The visual survey carried out in June of 2008 confirmed the presence of typical interveinal chlorosis symptoms in other greenhouses, requiring further studies to assess the incidence of ToCV on tomato crops. References: (1) H. Delatte et al. Plant Pathol. 55:289, 2006. (2) D. Louro et al. Eur. J. Plant Pathol. 106:589, 2000. (3) L. Segev et al. Plant Dis. 88:1160, 2004.