New Disease Reports
New Disease Reports (2009) 20, 25.
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First report of ‘Candidatus Phytoplasma asteris’ (16Sr I group) associated with yellows and little leaf diseases of Hibiscus rosa-sinensis in India

Yamini Chaturvedi 1, Maneesha Singh 1, S.K. Snehi 2, S.K. Raj 2 and G.P. Rao 1*

*gprao_gor@rediffmail.com

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Accepted: 16 Dec 2009

Hibiscus rosa-sinensis (family Malvaceae) is an ornamental perennial shrub and native of warm temperate, subtropical and tropical regions. It is widely used for pain treatment related to menstruation, cystitis, venereal disease, feverish illnesses, bronchial catarrh and cough (Chopra et al., 1986). Yellows and little leaf symptoms were observed on H. rosa-sinensis plants growing in different gardens and nurseries of Gorakhpur , Eastern U.P., India , during June 2008. Hibiscusisolate GKP-1 (from Betiahata garden, Gorakhpur) showed excessive yellowing, shortening of leaves and vein banding symptoms (Fig. 1a), while isolate GKP-2 (from Parijat nursery, Gorakhpur ) exhibited little leaf, curling, puckering and stunting of the entire plant (Fig. 1b).

Leaf samples from plants with and without symptopms were collected and total DNA extracted from 100 mg using a phytoplasma enrichment procedure (Ahrens & Seemuller, 1992). Direct and nested PCR for Hibiscus phytoplasma 16S rRNA were performed using P1/P7 (Deng & Hiruki, 1991) and R16F2n/R2 primers (Gundersen & Lee, 1996), respectively. A nested amplicon of expected size (approx. 1250 bp) was obtained for both GKP isolates, but not for symptomless plants. Amplicons were purified using a PCR clean-up system (Promega, USA) and sequenced (Genei Pvt. Ltd, India ); data were deposited in GenBank for GKP-1 (Accession No. FJ939287) and GKP-2 (FJ939288). BLAST analysis showed a 98% 16S rRNA sequence identity between GKP-1 and GPK-2, which suggest that these are two different 16SrI-related isolates that may be associated with different symptom manifestations in the same host. GKP-1 and GKP-2 exhibited 99% 16S rRNA sequence identity with those of members of ‘Candidatus Phytoplasma asteris’ (16SrI group), which was confirmed by phylogenetic analysis (MEGA 4.0).

Hibiscus sp.has been reported as a host for ‘Ca. Phytoplasma brasiliense’ (16SrXV group) in Brazil associated with a witches’-broom disease (Montano et al., 2001). However there are no reports as a host for ‘Ca. Phytoplasma asteris’-related strains. To our knowledge this is the first report of a ‘Ca. Phytoplasma asteris’ isolate associated with yellows and little leaf disease of H. rosa-sinensis.

Figure1+
Figure 1: Symptoms observed in H. rosa-sinensis with two isolates of ‘Ca. Phytoplasma asteris’: (a) GKP-1 (from Betiahata garden, Gorakhpur), and (b) GKP-2 (from Parijat nursery, Gorakhpur)
Figure 1: Symptoms observed in H. rosa-sinensis with two isolates of ‘Ca. Phytoplasma asteris’: (a) GKP-1 (from Betiahata garden, Gorakhpur), and (b) GKP-2 (from Parijat nursery, Gorakhpur)

References

  1. Ahrens U, Seemüller E, 1992. Detection of DNA of plant pathogenic mycoplasma-like organism by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82, 828-832.
  2. Chopra RN, Nayar SL, Chopra IC, 1986. Supplement to Glossary of Indian Medicinal Plants. New Delhi, India : Council of Scientific and Industrial Research.
  3. Deng S, Hiruki C, 1991. Amplification of 16S rRNA genes from culturable and nonculturable mollicutes. Journal of Microbiological Methods 14, 53-61.
  4. Gundersen DE , Lee I-M, 1996. Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopathologia Mediterranea 35, 144-151.
  5. Montano HG, Davis RE, Dally EL, Hogenhout S, Pimentel JP, Brioso PST, 2001. 'Candidatus Phytoplasma brasiliense', a new phytoplasma taxon associated with hibiscus witches' broom disease. International Journal of Systematic and Evolutionary Bacteriology 51, 1109-1118.
This report was formally published in Plant Pathology

©2009 The Authors