New Disease Reports (2009) 20, 16.

Definitive identification of orange rust of sugarcane caused by Puccinia kuehnii in Cuba

L. Pérez-Vicente1*, E.L. Martín-Triana2, F. Barroso3, E. Martínez-de la Parte1, O. Borrás-Hidalgo4 and I. Hernández Estévez4

1 Central Plant Quarantine Laboratory, National Plant Quarantine Centre, Ministry of Agriculture,  Ayuntamiento 231 e/ San Pedro y Lombillo, Plaza, C. Habana, Cuba
2 Villa Clara Provincial Plant Protection Laboratory, Ministry of Agriculture, Cuba
3 National Sugarcane Research Institute, Ministry of Sugar Industry, Cuba
4 Center of Genetic Engineering and Biotechnology, State Council of Cuba

*lperezvicente@live.com

Accepted: 28 Oct 2009

In November 2008, leaf lesions, pale yellow to yellow-orange, sometimes coalescing in large necrotic areas (Fig. 1), were observed on hybrid sugarcane clones (Saccharum spp.) CSG 86-504, CSG 24-92, CSG 204-92 and CC 87-409 at the Sugarcane Research Institute (INICA) Station in Villa Clara province, in central Cuba. Specimens were submitted to the Central Plant Quarantine Laboratory for identification. The observed morphological features were consistent with those described for Puccinia kuehnii (Virtudazo et al., 2001; Comstock et al., 2008). Uredinial lesions, variable in size (3-4 mm in length), were generally light brown in colour, but occasionally yellow-orange to cinnamon brown (Fig. 2). Hypophyllous uredinia were ellipsoidal to fusiform in shape, distinctly lighter and less uniformly distributed than for P. melanocephala. Urediniospores (Fig. 3) were mostly obovoid to pyriform, almost ellipsoidal in shape, variable in size, 27.5-55.0 (41.9) x 17.5-37.5 (26.0) μm, echinulate with evenly distributed spines (Fig. 4A and B). Walls (orange to light cinnamon brown in colour) were 1-2.5 μm thick, often with a pronounced apical thickening, 3-7 μm (mode 5 μm). Four to five equatorial pores were present. In contrast to P. melanocephala, paraphyses were not visible in stereoscopic views of the uredinia and were absent in most specimens. Telia and teliospores were not observed.

Total DNA isolation from symptom-bearing leaves with uredinia was performed according to Lin et al. (2001). A real time PCR was conducted using the SYBR green technique with primers synthesized from exact sequences of ITS1, 5.8S, and ITS2 of the large subunit rDNA region reported in GenBank Accessions EU164549 and EU176009 (Comstock et al., 2008). Results confirmed the identification as P. kuehnii. Previous reports of P. kuehnii in Cuba (CMI, 1969) were based on misidentifications of P. melanocephala. To the authors’ knowledge, P. kuehnii has not been reported by any diagnostic laboratory in Cuba in the last 30 years.In addition, Cuba was not included in the latest P. kuehnii distribution map (CAB International, 2007). Therefore, this is the first confirmed report of the presence of P. kuehnii in Cuba .The disease has since been found in different localities in the central and eastern regions of Cuba , without any reported economic losses to date.

Figure1
Figure 1: Lesions of orange rust on sugarcane leaves
Figure2
Figure 2: Uredinia of Puccinia kuehnii
Figure3
Figure 3: Young and mature Puccinia kuehnii urediniospores (Bar = 10 μm)
Figure4
Figure 4: Urediniospores of Puccinia kuehnii mounted in lactophenol cotton blue highlighting a) the thick wall apex and b) the evenly distributed echinulations (Bar = 10 μm)

References

CAB International, 2007. Crop Protection Compendium. Wallingford, UK : CAB International.

Commonwealth Mycological Institute (CMI), 1969. Puccinia kuehnii Butler . Distribution Maps of Plant Diseases, Map 215 (3rd edition).

Comstock JC, Sood SG, Glynn NC, Shine JM Jr, McKemy JM, Castlebury LA, 2008. First report of Puccinia kuehnii, causal agent of orange rust of sugarcane, in the United States and western hemisphere. Plant Disease 92, 175.

Lin R-C, Ding Z-S, Li L-B, Kuang T-Y, 2001. A rapid and efficient DNA minipreparation suitable for screening transgenic plants. Plant Molecular Biology Reporter 19, 379a-379e.

Virtudazo EV, Nojima H, Kakishima M, 2001. Taxonomy of Puccinia species causing rust disease on sugarcane. Mycoscience 42, 167-175.

©2009 The Authors